Biochemistry - Science topic

Hello,

My lab recently acquired a combination ORP electrode, with the refrence being Ag/AgCl and the working electrode being platinum.

We are hoping to use this electrode to measure the reduction potential of buffers prepared using biologically active compounds, such as GSH and GSSG and probe redox active systems by artificially setting the potential.

Recently, I was trying to validate the electrode by preparing 1 mM total concentration solutions of varying ratios of GSH and GSSG. This was done as, to my understanding regardless of concentration the ratio of Ox vs Red determines the potential value of the solution via the Nernst Equation. However the readings I got were all positive, and nowhere close to the expected potential, even when correcting for the electrode difference between Ag/AgCl and SHE.

Secondly, in 1x PBS pH 7.4, I added increasing amounts of BME up to 1 M and got an exponential decay like curve asymptotically approaching ~-120 (SHE) mV.

I am having trouble making sense of these results, namely the GSH vs GSSG ratio, and why the readings would not follow the nersnt equation.

Can anyone explain how to use these ORP electrodes, and where I may be going wrong in these experiments? All the information I can find online are referring to waste water treatment.

I am attempting to synthesize a schiff base ligand by reacting 5-hydroxytryptamine (HCl salt) and pyridoxal (HCl salt) in methanol, but the final product is not crystallizing in the methanol to be recrystallized. The product is originally a sticky brown oil that solidifies as a whole. What should I try differently? Sulfuric acid catalyst and maybe glacial acetic acid? Anything else?

Background Biochemistry

I tried glycine buffer but the yeast cells acidify the buffer so the pH goes down to 7-6 overnight. I need something that will stay around 9 and that isn't toxic to the cells.

Most of the researchers use to teach at university. In some careers, professionals who exert their profession without doing research share teaching spaces. When I was a chemistry student, 100% of my teachers were researchers ranging from PhD candidates to experts in their respective fields. While it may seem logical for researchers to be the best candidates to teach in fields such as chemistry or biology, what about healthcare-related fields like medicine, pharmacy, or biochemistry? Who is better suited to lead a class, a researcher or a professional, or both, each one in different subjects? We can distinguish between basic and clinical subjects. I am interested in hearing your thoughts on this matter.

In all academic sources, sucrose is identified as α−glucose (1-->2) β−fructose. However, I cannot find any explanation anywhere as to why the fructose monomer has to be in the β configuration. Maltose has both α and β anomers, same for lactose. Even trehalose, another non-reducing disaccharide with glycosidic linkage between two anomeric carbons, has α-α, α-β, and even β-β anomers. Why is sucrose special? And is there a disaccharide out there that has α−glucose (1-->2) α−fructose configuration?

A patient with desminopathy (mutation Thr341Pro DES in a heterozygous state) with the progression of the disease has a decrease in taste and smell, immunosuppression, and an increase in IgA in the blood.

Oddly enough, but all this is characteristic of infections, including viral ones. For example, it is known that if the hepatitis C virus is not treated, then death will occur in 20 years.

In the identified case of late onset desminopathy, muscle weakness manifests itself at the age of 30, and death occurs 20 years after the onset of the disease.

I need some suggestions for articles recently focused on the computational design of proteins, as well as the evaluation of the designed proteins by assessing various properties or values. If anyone is interested in this field and has read articles on this topic, please don't hesitate to share your suggestions below

Dear Researcher Gate,

I inquired about adding a reviewer to the submitted manuscript in Biochemistry can you help me with this?

Thanks

maha saad

I am currently learning about PyMol to utilize in my project. I used PyMol to visualize potential H-bond interactions in specific amino acid residues. However, I have discovered that Arg465 and Ser461 show a distinct interaction, as shown.

Please help identify this interaction.

An unopened Sigma-Aldrich (P4557) phenol solution bottle was shaken (prior to the addition of the Equilibration Buffer) and a gel-like layer formed at the bottom of the bottle. The upper phase is still liquid. The bottle was shaken briefly after the phenol solution was taken out of +4 C. What should be done? Should it be heated in order for it to return to liquid?

Hi,

I am looking for build up a consensus or a group in clinical biochemistry for mutual exchange in scientific idea, research protocols, cooperation in proposal preparation, sharing in books, Research articles and reviews.

It is known that in the early stages of desminopathy the muscles most often affected are: Semitendinosus, Gracilis and Sartorius. What is the reason for the damage to these particular muscles?

Here and Now: Dependent Clause (Truth: Millions of Colors, Shades of Grey)

Here and Hereafter: Independent Reasoning (Independence: School of Taught)

Hereafter and Evermore: Organized Biologics (Chemistry x Biology + Physics) = Location Location Location = L³ (Heart-Person, Soul-Public, Interest-Private)

Key: Spelling-PEAR$

Note: Commentary

Comment: oh holy nigh

In biochemistry, most work is done at room temperature. Yet, basic thermodynamics tells us that affinities often change with temperature, in positive or negative directions depending on the entropy/enthalpy contributions. Thermal transitions can occur which both quantitatively and qualitatively change the behavior of the molecules of study. In enzymatic reactions rate-limited by diffusion-mediated product release, the increased rate of diffusion could increase the rate of product release above the rate of the chemical step, such that the chemical step becomes rate-limiting. If one discovers a compound that potently inhibits this enzyme at 25C, it may have reduced effect at 37C, or none at all. Likewise, if an enzyme is predominantly dimerized at 25C based mostly on enthalpic contributions, this dimer may not even exist at 37C. Screening compounds against the dimer may be of little relevance to the situation in vivo. The converse could happen if dimerization is entropically driven. Temperature-dependent changes in solution properties can also obscure the relevance of 25C results to 37C, such as viscosity.

I welcome everyone's two cents.

Here is an excerpt from Kollmann et al. (2020, J. Physiol.):

"This ring was placed in a recording chamber continuously perfused with 37°C aerated Hepes solution containing (in mM) 136 NaCl, 10 glucose, 5 KCl, 10 Hepes, 1.2 MgCl₂, 2.5 CaCl₂ (pH 7.40) at a rate of 11 ml min^-1. Hypoosmolality of the Hepes solution was established by reduction of the NaCl content to 33 mM (94 mOsm kg^-1 H2O), 58 mM (144 mOsm kg^-1 H2O) or 83 mM (194 mOsm kg^-1 H2O)."

For validation, I tried to calculate the osmolality by myself. For example, when the NaCl content is changed to 33 mM, the osmolality should be

33*2 (NaCl)+10*1 (Glucose)+5*2 (KCl)+10*1 (Hepes)+1.2*3(MgCl₂)+2.5*3(CaCl₂)

which gives 107.1 mOsm/L, i.e., 107.1 mOsm/kg. But 107.1 is largely different from what the author stated, namely 94 mOsm/kg.

When the NaCl content is changed to 58 mM or 83 mM, the osmolality should increase by (58-33)*2=50 mM, or (83-33)*2=100 mM. This is consistent with the author's calculation, i.e., 144-94=50 mM, or 194-94=100 mM. So, my calculation is correct at least in terms of NaCl. But how can I calculate the contributions of the remaining solutes correctly?

Reference: Kollmann, P. et al. Submucosal enteric neurons of the cavine distal colon are sensitive to hypoosmolar stimuli. J. Physiol. 598, 5317–5332 (2020).

Is it correct to filter the extracts we obtain with Whatman filter paper and use them directly in experiments to investigate the bioactivity of phytochemical substances? Or is it correct to first centrifuge the extracts we obtain, then filter the supernatant and use them in experiments?

Some (but not all) DNA polymerases such as Klenow fragment, Taq, and Phi 29 DNA polymerase can catalyze strand displacement synthesis. This is evidenced by opening of molecular beacons and Loop Mediated Isothermal amplification (LAMP).

in strand displacement synthesis, the primer strand is extended using the template strand and each nucleotide incorporated displaces the 3rd strand that was originally annealed to the template?

where is the third DNA strand? the one being displaced? crystal structures and cryo-EM structures of linear primer template dsDNA structures with magnesium and dNTPs and DNA polymerases are not informative about where 3rd strand of DNA is. The displaced strand is not the template and is not the primer.

Where is the displaced 3rd strand of DNA?

I need to dissolve sulphate to purify the nanozyme synthesized with magnesium sulphate

Dear all,

I have been trying to knockdown my target protein using siRNA. The protein has several isoforms the effect of this knockdown has been showing up in the functional assay. However, I don't see the knockdown in the wesptern blot of the same test sample. Please suggest how to do an effective knockdown so that I can visualize that in the western blot as well.

Thank you in advance for your kind suggestions

Sincerely,

Prem

Hello,

We are trying to purify proteins using a secretion system but do not have TFF cartridges for our device. Does anyone know where we can purchase these? Or is there a better replacement? We want to downscale the volume from 3 L to 100-200 mL.

I've attached a picture for reference.

Thanks,

Thomas Newton

I'm on the lookout for remote bioinformatics and computational biology opportunities where I can actively contribute to research projects. Compensation is not a priority for me; my main focus is to gain hands-on experience in these fields.

#biopython

#computational_biology

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#R

I intend to explore the integration of computational biology and artificial intelligence (AI) with laboratory and experimental work, encompassing animal models, cell culture, clinical trials, and molecular studies. As a clinical biochemistry student with a keen interest in AI, I believe this interdisciplinary approach holds immense potential for advancement and innovation.

However, I face the challenge of identifying relevant literature in this emerging field. I would greatly appreciate guidance on effective keywords and search strategies to navigate this landscape of research and achieve my research goals.

My target protein is a membrane protein and I want to check its expression level in the test sample using western blot. Can anyone suggest which protein should be used for loading control, like we use beta-actin in cytosolic proteins. And is there antibody available for that ?

I intend to explore the integration of computational biology and artificial intelligence (AI) with laboratory and experimental work, encompassing animal models, cell culture, clinical trials, and molecular studies. As a clinical biochemistry student with a keen interest in AI, I believe this interdisciplinary approach holds immense potential for advancement and innovation.

However, I face the challenge of identifying relevant literature in this emerging field. I would greatly appreciate guidance on effective keywords and search strategies to navigate this landscape of research and achieve my research goals.

I need to convert from micromoles per second per liter (µmol s⁻¹ L⁻¹) to millimoles per gram of dry cell weight per hour (mmol gDCW-1h-1)

Thanks!

Hello everyone! I am new to this platform, I am looking for this book: "Bioquimica del estres Oxidativo" (Diego Camps, 2010)

I would like to know if anyone has it in digital format to share it with me. Waiting for a reply. Thank you very much!

I need to extract the liver from frozen Emerald rockcod and sequence its RNA. To keep the RNA stable and prevent degradation, I would like to avoid thawing the fish for the dissection. However, I haven't been able to find any methods that keep the fish frozen. Does anyone have any tips on how to best achieve this?

If an active site mutant knocks out product formation at all excessive concentrations of substrate and at all excessive concentrations enzyme, but substrate binding affinity via anisotropy shows no difference in binding affinity for wildtype versus active site mutant enzyme, is kcat = 0? But if kcat = 0, then by the relationship of Michaelis constant (K_M) to kcat then K_M = K_D.

How do you show to reviewers that the active site mutant is dead? If the wildtype mutant starts producing product in seconds, are you supposed to measure the reaction for the mutant for an hour at zero concentration of substrate and at 100fold excess substrate concentration of the K_M for the wildtype enzyme for the active site mutant, and show the time course?

Are there any publications that show a mutant enzyme is not just slow to produce product but is rather incapable of producing product but can still bind substrate? Examples would be greatly appreciated!

Hello all,

I hope everyone is doing well.

We are preparing buttermilk from standardized milk. In this regard we need to increase the pH of the buttermilk. We used sodium citrate (0.4% & 4%) and sorbitol (0.4%) to check any change in the pH. Unfortunately, there were no significant changes observed.

Please suggest any recommendation to increase pH using any natural or food-grade chemical compounds and any other alternative options.

Thank you in advance.

Keep smiling and Stay healthy.

I am reaching out to #researchers in the field of #Biochemistry, #Biophysics and #Bioinformatics, for collaborative partnership in scientific research. The researcher should be academic staff at the tertiary institutions in following listed countries:

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Interested researcher should kindly email to [email protected] with the subject: Research Collaboration from "your country".

Thanks.

Toluwase H. Fatoki

Visionary @ Heze-Sapience International, Nigeria.

Lecturer @ Department of Biochemistry, Federal University Oye-Ekiti, Nigeria.

Announcement link: https://www.linkedin.com/posts/hezekiahfatoki_researchers-biochemistry-biophysics-activity-7115426383475990528-lr6X?utm_source=share&utm_medium=member_android

Is there any system with small molecule binders and a short protein tag that is higher affinity than 6xHIS-NTA?

Dear all, please suggest or provide a link for the provider or company who can design or synthesize shRNA plasmid (Single vector system) that will directly express into the mammalian cells to knockdown the protein expression for long term. Any suggestions will be highly appreciated.

Thank you

with kind regards

Prem

Fields:

Cell and Molecular Biology

Biochemistry

Physiology

Microbiology

I am trying to express several proteins at the same time, but I want to use a different promoter and terminator for each one to avoid the possibility of recombination.

The promoters that are available to me are: TEF2, PGK1, CCW2, TDH3 and HHF2. The available terminators are: ENO1, SSA1, ADH1, PGK1 and ENO2.

Has anyone ever used these combinations of promoters and terminators? In your experience, which combinations work the best?

Hi

I'm purifying some mutants of the protein I study. The wild type protein exists as a monomer and is 28kDa.

I have two mutants (same protein, same number of amino acids but with 8 amino acid substitutions at defined positions), one of the mutants (mutant 1) analysed using size exclusion chromatography with multi-angle static light scattering (SEC-MALS) and its MW was shown to be 33kDa and has an oligomerization state of 1.2. The other mutant, mutant 2, also measured by SEC-MALS was 59kDa with an oligomerization state of 2.2.

For the wild type to measure the concentration I've just been using the MW (28kDa) and extinction coefficient (calculated by entering the sequence into online software ProtParam) and using a NanoDrop measuring absorbance at 280. This gives the concentration in mg/ml which I then convert to molar concentration.

For the mutants I want to measure their concentration the same way - measuring A280 on the NanoDrop using the mutants MW and extinction coefficient and calculating molarity from mg/ml. I'm not sure if this is an obvious/stupid question but what MW weight and extinction coefficient would you use for the mutants on the NanoDrop? E.g. For example mutant 2 molecular weight (MW) of the protein based on its amino acids (AA) composition is predicted to be 28kDa, but SEC-MALS shows it is 59kDa as the protein forms an oligomer.

My instant is to use 59kDa and the computed extinction coefficient predicted from the AA composition - is this correct?

Thanks in advance!

Dear All,

Ph.D. full-time position in Bangalore with fellowship:

Eligibility: M.Sc. Chemistry/Biochemistry/Biotechnology/Microbiology/Bioinformatics with first class of 60%.

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PhD biochemistry candidate is required for the post of guest faculty. Interested candidates can contact me asap.

I learned DNS assay to find out the quantity of maltose in the sample.

I realized the importance about the boiling water in this assay. But I got a several question after that.

I don't know the reason why do I have to cool-down the sample after heating up?

And also is ice a tool used to reduce cooling time, or is it used to cool quickly?

Please leave me a response about this problem.

Thank you for your help.

Joonseo_Cha The Student of Hallym University

How many grams of K2Cr2O7 to dissolve it in 1 liter Distilled water to obtain 50 ppm of Chromium? to become aqueous solution, Is there a specific equation to apply? Thanks

Ali

The size differ little bit from one species to another, yet they have one size range. Also, the size of them in their native form so they don't lose their colour while isolation.

I saw these on a same step of conversion of Malonyl CoA to Malonyl-ACP. Could anyone clarify this? I add that reference link below.

KEGG PATHWAY: Fatty acid biosynthesis - Chlorella variabilis (genome.jp)

I'd like to perform a SEC step prior to on-column refolding of a protein I am expressing/purifying but am worried about the extended time the protein will be in the presence of urea (and subsequent protein carbamylation) in the current refolding workflow I have.

Is there any concern of protein modification with a 4-6 hour denatured protein purification workflow using 6M GndHCl?

I am a second year PhD student at Wayne State University looking to screen a library of photosensitizers (transition metal complexes) for activity with ultrasound irradiation in cell cultures (likely MCF-7A cells). I would ideally like to set this up in a 96-well plate to screen multiple compounds but I am unsure of the set up as previous literature has not been as helpful as I would hope. The instrument I have is 3 MHz with a 3.5 cm diameter probe (Model-Sonic 103, Preamonex). I am unfamiliar with soundwaves so I will briefly discuss what I know so far from previous literature.

Overall, 0.3 W cm–2, 3.0 MHz seems to be a fairly common treatment condition when combining compounds with cell cultures.

This source clusters samples in a 2x2 square in the well plate and places the probe underneath the plate per cluster (see SI). Does a coupling agent need to be used and can this be done while keeping the plate sterile for continued incubation? Do the ultrasound waves pass through the plate to other clusters of wells (in a way that would significantly affect the other clusters)? Is there a way to evenly apply ultrasound stimulation evenly across the whole plate?

This source works with nanoparticles and a 96-well plate, but details of the set up are not given. Unless I'm missing something?

This source appears to place a sponge in degassed water on the head of the 35 mm probe. Could this be a valid option for the cluster of wells (2x2 square) mentioned above?

For the purpose of transfection, a 6-well plate was put into an ultrasound water bath.

Ideally, I would like to use the probe (as opposed to a water bath) to keep the instrumentation consistent between in vitro and in vivo studies. As I mentioned, I'm very unfamiliar with sonication/ultrasound and its physics. Any assistance/advice on how to make an accurate and precise high throughput set up is appreciated.

Thank you in advance!

How often do you get new salts? What salts do you get "fresh" most frequently? All help is greatly appreciated. Thank you to everyone who reads or responds, hope you all have a fantastic day.

I am a Graduate Student at the University of Wisconsin Milwaukee. Our most recent electrophysiology experiment was going fantastic for about 3 weeks. Suddenly, every slice was very poor quality, and every cell we patched onto died within about two minutes. Even if you don't have an Electrophysiology background, all advice and input is appreciated. It's been two weeks since this problem first arose. We didn't alter any procedures, everything has been held constant. We don't know what could be causing the sudden change in slice quality.

So far we have three theories: DDH20 filter needs replacing, our glassware has somehow become contaminated, or our salts have gone bad.

The DDH2O water resistance reads about 14.8MOhms. We changed the filter about a year ago. From what I have read, DDH2O should be between 14 and 18 MOhms to ensure slice quality, so we think our water is fine.

Our glassware washing procedure is 3 rinses of tap water, 3 rinses of deionized water, and 1 rinse of DDH2O. We don't know how else we can safely clean the glassware for slice preparation. But we don't think this is the issue.

Our main theory is the salts have gone bad. Our lab is on the 4th floor, and it is usually quite warm and humid on our floor during the summer. Most of opened salt bottles we use were first opened between 2 and 7 years ago. (for example, our bottle of sodium phosphate monobasic monohydrate and our potassium chloride were both opened 6 years ago. Our magnesium chloride and sodium chloride were both opened 2 years ago)

We think with repeated opening of the salt bottles causes debris/water from the atmosphere to leach into the salts, resulting in a change in our solutions and causing the cell death we have been observing. Is this a real possibility? How often do you get new salts? What salts do you get "fresh" most frequently?

We checked the pH, the pH of our solutions is within .1 of what it should be. All help is greatly appreciated. Thank you to everyone who reads or responds, hope you all have a fantastic day.

I am currently making bacmid and consequent baculovirus using pDEST8, pFastBac and 438a vectors and recently switched to DH10EmBacY cells instead of DH10Bac due to the added YFP signal to monitor transfection, etc.

I was wondering if there is any reason for me to not use either of the two competent cells interchangeably to make bacmid DNA?

I observed an interesting pattern that resembles a "flattening" of the near-UV region of the absorbance spectrum of the rat intestinal mucus in the streptozotocin-induced model of sporadic Alzheimer's disease. I'm studying qualitative and quantitative alterations in mucus as there seem to be some changes in the gut-brain axis, intestinal redox homeostasis, and cell turnover ( , ). I observed a quite dramatic change in the number and responsiveness of goblet cells so the observed spectral shift may be driven by a component of mucus. Another possibility may be a component of bile, a peptide or bilirubin/biliverdin, or even complexation between bilirubin/biliverdin and some other molecule (e.g. I found a similar pattern in a publication by Klinke et al. who reported flattening of the biliverdin near-UV spectrum upon binding to bacteriophytochrome? ). Any ideas about what I may be looking at?

Thanks!

Dear community,

I am planning on conducting experiments for which I need to obtain a plasma-free platelet suspension from an aliquote of a platelet concentrate. Do you now any methodology/protocol that allows for washing without extensive cell activation? I need the cells to be a "functional" as possible.

Yours sincerely,

Michael

I would like to add BG-azide to cells for SNAP tag pulldown however I don't know whether it is going to be cell permeable. My instinct is to day yes it will be but neither me nor the supplier know whether that is true. I thought I would ask if anyone has tried something similar, even though that is unlikely... I have attached the structures in case that is informative

These are the products https://www.iris-biotech.de/global/rl-3950 and https://www.iris-biotech.de/global/rl-3960

In a patient with desminopathy (mutation Thr341Pro DES in the heterozygous state) with the progression of the disease, we note signs and symptoms that are also characteristic of botulism: bradycardia, arrhythmia, AV blockade, a significant decrease in the average duration of motor unit potentials according to electroneuromyography, paresis and paralysis of the striated muscles, decreased sweating, paresis of the gastrointestinal tract, dry eyes, dry mouth, symmetry of neurological symptoms, hoarseness, impaired visual acuity, doubling of objects occurs, progressive muscle weakness. These signs and symptoms are characteristic of botulism, only when a case of desminopathy is detected, they proceed slowly.

Hi everyone.

I have protein with concentration of 0.6 mg/mL

The total volume of my protein is 4 mL.

Protein size is ~18 kDa

How can I convert my total protein concentration to micro molar?

Thank you.

There seems to be some consensus on the reaction mix for lactate dehydrogenase (LDH) activity assays, but the composition of the homogenization buffer (HB) differs wildly among studies. Maybe somebody can help me with the following three questions:

Any recommendations and/or tips regarding LDH assays are greatly appreciated. Thanks!

Studies polled for this question:

Chippari-Gomes AR, Gomes LC, Lopes NP, Val AL, Almeida-Val VMF (2005) Comp Biochem Physiol B Biochem Mol Biol 141:347–355. https://doi.org/10.1016/j.cbpc.2005.04.006

Driedzic WR, Fonseca de Almeida-Val VM (1996) J. Exp. Zool. 274:327–333. https://doi.org/10.1002/(SICI)1097-010X(19960415)274:6<327:AID-JEZ1>3.0.CO;2-Q

Gerringer ME, Yancey PH, Tikhonova OV, Vavilov NE, Zgoda VG, Davydov DR (2020) FEBS J 287:5394–5410. https://doi.org/10.1111/febs.15317

Heinrichs-Caldas W, Almeida-Val VMF de (2021) Fish Physiol Biochem 47:1759–1775. https://doi.org/10.1007/s10695-021-01000-0

Martinez ML, Chapman LJ, Grady JM, Rees BB (2004) J Fish Biol 65:1056–1069

Moon TW, Mommsen TP (1987) J. Exp. Zool. 244:9–15. https://doi.org/10.1002/jez.1402440103

Sidell BD, Driedzic WR, Stowe DB, Johnston IA (1987) Physiol Zool 60:221–232. https://doi.org/10.1086/physzool.60.2.30158646

Singer TD, Ballantyne JS (1989) J. Exp. Zool. 251:355–360. https://doi.org/10.1002/jez.1402510312

Hello everyone!

I recently received invitations to contribute an article to the current issue of the Journal of Brilliant Engineering (BEN) and Chemistry and Biochemistry. Can anyone with experience or knowledge about these journals confirm their originality?

Thank you.

Hey all, I'm very interested in understanding more about the biochemistry behind all of the wash buffers that go into the streptavidin affinity purification protocol seen here (https://static1.squarespace.com/static/5617d7d8e4b09f2fdf34baa6/t/5fe2631ba775596e136098b6/1608672031257/Cho+NP+2020.pdf pg 3990, step #38).

I understand this is to help reduce non-specific interactors from being pulled down with the biotinylated proteins; however, I'm more curious to understand the necessity of the 2M Urea vs sodium carbonate vs KCl, etc....looks like previous iterations of this protocol have not used sodium carbonate and KCl buffers but I can't find substantiation for why they were added.

Thanks in advance!

biofilm and quorum sensing genes of E.coli, not used drug but chemical material (Thiophene)

I basically have normalized fluorescence data generated from a series of protein dilutions. I need to calculate K, K should be the same in all dilutions, thus the need for a global fitting approach. I have been using XLfit, but would like to move into a script format. Thanks in advance! Thus far, I found the package renz, but to my understanding, it does not accomodate for global fitting. Thanks in advance!

Specifically interested in purine derivative as a cation of ionic liquid.

Any relevant reading suggestions are highly appreciated. Thanks.

1-The conditions of the relationship between the pigment (Pyocyanin) with Gold (Au Nanoparticles)?

2-Dose the pigment (Pyocyanin) work as Reducing agent or not?

3-Dose the pigment (Pyocyanin) endures high temperatures or not?

One example would be MBP and MBP-74, an interaction which can be disrupted by maltose when it binds MBP.

Hansen Solubility Parameters full sets (dD, dP, dH and Ro) for

1) Cholesterol CAS#[57-88-5], and/or

2)Cholesterol Monohydrate with CAS#[5808-12-8] are needed.

Any or both experimental or calculated in HSPiP values are interesting.

Old published values (20.4, 2.8, 9.4 and 12.6) are off interest.

I am currently working on Co-IP of NRIP and MyoD by overexpressing both proteins in HEK293T cell. The question is I cannot get a specific band of MyoD. For my input, I think the protein level is too low to be detected so I will increase the protein amount to run a western blot. However, several bands appear when detected using MyoD antibody after co-IP. The band most likely MyoD is pointed with a blue arrow. Some of my colleagues say it may be endogenous MyoD or protein with similar conformation. Does anyone face a similar situation when working with this protein? or any reference paper suggested for studying this protein. Thank you.

Can alpha glucosidase and alpha amylase work at room temperature and at 37*C? If so why? The specifications for these enzymes are that they work at 37*C. But number of papers have modified their protocols 25*C. How can it work at both room temperature and at 37*C?

Hello

I'm sure this question has been asked a lot but the protein I am purifying is not as clean as I would like and all the potential solutions I have read about have not worked for me.

I am purifying a protein that forms inclusion bodies. The construct is 14x His tag - Tev cleavage - protein of interest. I use BL21 Star (DE3) cells and TB media for growing the bacteria and induce at OD 0.6 for 4 hours at 30 degrees.The purification protocol is based on a previously published paper. Following sonication, lysis buffer and washing the pellet the protein is extracted from the inclusion bodies using 6M Guanidine Hydrochloride, applied o/n at room temperature to a Ni NTA agarose column. It is then eluted in 4M Guanidine. I then pass it through a RP-HPLC. As you can see by the attached image although I am getting a high amount of protein it has a large smear which I am not sure if it is contaminants or degredation. I overloading with sample when trying to judge purity but I think it's better to get an accurate representation of what’s going on rather than kid myself I am working with a pure sample.

The HPLC makes no difference so I think optimisation on the Ni-NTA purification is needed but nothing thus far has worked, I have tested different induction temperatures, leaving it on the Ni column at both 4 degrees and for a few hours (rather than O/N at RT like the protocol says), protease inhibitors and washes/step-wise elutions with different concentrations of imidazole (as in attached file) - yet none of this has worked. I've even thought about making the His tag smaller (14x seems quite large, but I cant see how this would impact purity other than needing more imidazole for elution).

If anyone has comments on the purity and has any tips that I have not tried it would be greatly appreciated!

Thanks!

In speculating about plant migration from the oceans to land, I wondered if any plant species went back into the ocean. Since that is unlikely, this question arose to explain why.

In a study of plant genetics, this area of plant diversification is likely amazing, because of how the gene sequences are accessed and how complexity developed as new genes and biological systems.

Hello every body I want to assay lipase but I cant dissolve PNPP in isopropanol could you please help me in this way?

We are doing a research on Biofilm formation of Bacteria, for knowing each isolate strong, moderate or weak, by calculations tables of Standard deviation, Variance and Cutoff (Ct) etc… and with the help of Microsoft Excel but the results in the program differ from the hand written and don’t know the best way to calculate and compare the results, any help will be very appreciated, Thank you

Ali

In the Column treatment of aqueous solution of Chromium Removal by Adsorption

Bio-sensor development demands a variety of fields in cooperation. However, this breakthrough would not result in the case of doing research with a single attitude. I am calling researchers from biology or biochemistry, genetics, or related science to come into the discussion. Let me know if somebody has any knowledge in this field.

#Bio-sensors #Biochemistry #Genetics

My aim is to detect nascent protein synthesis by click-chemistry of L-homopropargyl-glycine-labelled proteins with biotin azide and detection by western blotting with fluorescently-conjugated streptavidin. So far, I have failed to detect labelling above what looks like background biotinylation. I'm using biotin azide from Biotium (Cat# 92167), but I've noticed there different biotin azide molecules with different molecular weights. I was wondering if there were differences in their applicability for copper-catalysed click-chemistry. Any help with troubleshooting this would be appreciated. Cheers!

Aside from inhibiting protease activity (not effectively working), I read some papers about doing modifications to the antibodies:

using unnatural amino Acids

using mPEG

But, are there better options, more commercially applicable?

Please spread the word: Folding at Home (https://foldingathome.org/) is an extremely powerful supercomputer composed of thousands of home computers around the world. It tries to simulate protein folding to Fight diseases. We can increase its power even further by simply running its small program on our computers and donating the spare (already unused and wasted) capacity of our computers to their supercomputation.

After all, a great part of our work (which is surfing the web, writing texts and stuff, communicating, etc.) never needs more than a tiny percent of the huge capacity of our modern CPUs and GPUs. So it would be very helpful if we could donate the rest of their capacity [that is currently going to waste] to such "distributed supercomputer" projects and help find cures for diseases.

The program runs at a very low priority in the background and uses some of the capacity of our computers. By default, it is set to use the least amount of EXCESS (already wasted) computational power. It is very easy to use. But if someone is interested in tweaking it, it can be configured too via both simple and advanced modes. For example, the program can be set to run only when the computer is idle (as the default mode) or even while working. It can be configured to work intensively or very mildly (as the default mode). The CPU or GPU can each be disabled or set to work only when the operating system is idle, independent of the other.

Please spread the word; for example, start by sharing this very post with your contacts.

Also give them feedback and suggestions to improve their software. Or directly contribute to their project.

Folding at Home: https://foldingathome.org/

Folding at Home's Forum: https://foldingforum.org/index.php

Folding at Home's GitHub: https://github.com/FoldingAtHome

Additionally, see other distributed supercomputers used for fighting disease:

Rosetta at Home: https://boinc.bakerlab.org/

GPUGRID: https://www.gpugrid.net/

When staining with hematoxylin and eosin of a muscle biopsy from a patient with T341P desminopathy, we observe accumulations of inclusions similar to nuclei (arrows in figures 1 and 2, x280). And outside of these accumulations - adipose tissue, which used to be muscle tissue. There are no such massive accumulations of inclusions in adjacent muscle fibers. We assume that clusters of inclusions are not nuclei? Figure 2 is the inverted figure 1.

For the protein and ligand preparation process what kind of structure should ı choose and what is the reason of that

We have a Flask that contains broth, and we want to inoculate it with Bacteria inoculum, Can we simply take a touch by the loop or by micropipette?