Biochemistry and Molecular Biology

Hi, I'm currently struggling with deriving the Adair equation for representing substrate binding cooperativity. So far, I have: p -> PS (reversible reaction, rate of k1) and PS -> P2.S (reversible reaction, rate of k2). y = number of unbound sites / total number of binding sites y = PS + 2P2.S / 2(P + PS + 2 P2.S) I have rate equations for the 2 reactions as k1 = PS / (P * S) k2 = P2.S / (PS * S) From this, I got PS = k1 * P * S P2.S = k2 * PS * S From this, I inserted PS into the rate equation for P2.S so I got: P2.S = k2 * (k1 * P * S) * S I multiplied out the brackets to get k2.k1 + k2.P + k2.S^2. I know tha…

Whats your opinions about the HSCs(Hematopoitc Stem Cells) in the treatments of Blood Disorders?? According to me, there are hopes, and HSCs will be the only way to treat all blood disorders even blood cancer.

Again, trying to do research into addictions... How exactly does the dopamine circuit work and how does it work in addictions (e.g. cigarette, or even video game addictions) And would taking in dopamine medication lead to a person craving cigarettes more? Could a person take dopamine before, let's say, practicing piano and become addicted to piano? Or would he enjoy the activity of piano more? If you lack dopamine while you perform a certain activity (let's say I force myself to play games even though I don't feel any urge to do so) does my addiction to the games weaken? Thanks!

I am working on Lambda Phage project. . AS a start, I should provide Lambda DNA marker/ EcoRI, Lambda/ Hind III and Lambda/ EcoRI+HindIII. I provided the phage from DSMZ the German company. We have transformed the phage in to the E.coli cells. We have got the lysogenic strains. Then we have tried to shift the bacteria into lytic phase and get the phage. 1 time it went properly and we could get good amount of DNA after purification by DEAE column, recommended in Current Protocol. When we digested the DNA with the given restriction enzymes, the pattern of Lambda/EcooRI was slightly different in one band and in the case of …

anyone can tell why .......Why carcinogens Bind to Nitrogen atom at N7 position and at O6 position of Guanine ?

Hi everyone! i am new to the whole biochemistry world and am attempting to extract collagen from arterial tissue. I have all the protocols in place and the techniques researched the only thing i am missing is the understanding as to what i am actually doing!! i was hoping somebody might be able to provide me with a link to some literature explaining how each type of soluble collage i.e. salt, acid and pepsin are actually extracted on a molecular level? Is it the bonds that are destroyed?? Also does any body know if there as a direct link between the differt types of soluble collagen and the mechanical properties of arterial tissue i.e. strength, stiffness etc…

Hi there, noob question, but i just looked a bit at cellular respiration and seeing that DHAP and PGAL are at the peak of the glycolysis energy curve wondered why, simply put, you could not eat those instead of glucose (cf. NADH). Are they toxic, instable, ...? What about other metabolic intermediates? Thanks!

This is a picture of a spike that shot up from my ice-cubes because the ice is 100% acutely distilled "empty" water. I thought it was facinating that the water was able to freez so uniformely because there are not any minerals in the water to hold the molecules down while the water expands. While the water was expanding it must have held a pocket of warm water then when it expanded to the point where the warmer water had no where to go the expanding forced a stream upwards and the stream froze in spike. If I could only get a camera in my freezer while this is happening.

Hi everybody I'm posting 2 links on articles written by a russian team doing research in genetics. The articles are quite opinionated but I'd just like to know what ppl think of what they are saying. Are they just talking jargon to cover up the fact they are wrong; if they are criticising mainstream views in biology, or is there some substance to their argument? The links are shown below, I would be very grateful for replies, as I placed a simliar post in the Physics section named Quantum evolution and haven't got many. http://www.emergentmind.org/gariaevI2.htm http://www.emergentmind.org/gariaevI3.htm Thanks in advance:-)

Is endocytosis a type of active transport? Shouldn't it be a type of an active process, but NOT active transport? My argument is as follows: Active transport by definition is; an energy consuming process that involves the movement of particles from a region of relatively low concentration to a region of relatively high concentration, AGAINST a concentration gradient. Endocytosis on the other hand does not necessarily involve the movement of particles against a concentration gradient. Instead, endocytosis is used to ingest particles by a cell which are too large to pass through the plasma membrane. For example: Consider a macrophage which has ingested two bact…

Over the past few months our lab has been attempting to measure expression differences between promoter sequences through transfection. We have a pGLOW plasmid to measure GFP expression on our two sample groups and a pCMV-LacZ control to measure transfection efficiency and number of plasmids per sample. The trouble we have run into is that GFP is expressed only if our LacZ plasmid is not present. We think something about the LacZ trasnfection interferes with pGLOWs transfection, and only LacZ gets transfected into the cells. I have a couple of questions: First: Has anyone come across this problem before? If so, how did you manage to fix it? Second: I find t…

In my naive understanding, single Celled organisms swim through their lives, "smelling / tasting" various chemical gradients, and thereby orienting themselves and their movement strategies. How do single Cells "taste" chemical gradients -- do they use chemo-receptors, on opposite sides of their membranes, to get a "forked-tongue" effect, and "smell in stereo" like snakes ? If you consider such chemo-receptors as "sense organs", what, inside the cell, is the "sensory data processor" -- to wit, the Cell's "proto-brain", which translates "taste" into movement instructions ?? If, in higher animals, the "seat of cognition" seems associated with the physical brain ("sen…

The last sentence in this wikipedia article: http://en.wikipedia.org/wiki/Diatom claims that "It has also been proposed that diatoms could be used as a component of solar cells, by substituting photosensitive titanium dioxide for the silicon dioxide normally used in the creation of cell walls." However, I could not open the reference EE article it cites. Does anyone know if this is actually possible? which approach would be the ideal one?

Hey, Having an issue with 293T cells and FGF stiumaltion assays. I grow 293T cells, transfect them with expression vectors containing proteins that i know increase response to FGF stimulation compared to untransfected cells, and i know that 293T cells endogenously express fgf receptor. i know the expression vectors express protein, via western blot, but the response to FGF stimulation after incubation in minimal media (krebs HEPES buffer) does not follow the expected outcome. when comparing even untransfected, non-fgf treated cells with fgf treated cells, no increase in pERK is seen (considered the measurable output for fgf stimulation), so the issue seems to be with…

Hello Everyone, I have been assigned to write a HYPOTHETICAL research proposal for a graduate level Biochemistry class. I have been racking my brain for possible topics, as well as scouring dozens of journals but I cannot seem to find any that will provide me with a working hypothesis that could utilize reasonable biochemical techniques. If you have come accross any recent topics/papers in your travels that have any unanswered questions that could be easily solved I would really appreciate it if you would post them below. Thank you!!! I just need a few ideas to maybe get me in the right track.

Hello All, We are facing a peculiar problem. I have attached my SDS PAGE gel for reference. We get a pattern as if protein has spilled into adjacent well and the blank well (1st and last) also shows band. we have been performing the same method for same protein for about 7 yrs with no difficulty but from past 2-3 months are encountering this problem. we have tried preparing fresh reagents and do the test but same results. what can be the problem as while loading the protein does not spill, still this results is confusing. Can anyone throw some light or elaborate on this. Any help will be appreciated. Thanks in anticipation

Hello, this is an unusual problem, but there may be someone out there who has been in a similar situation. I have an experiment where I induce hypoxia on cell samples, collecting them at specific time points over 24 hours, and measuring a particular connexin level in them. I measure the protein concentration of the samples using a Bradford Assay, to ensure that I have the same level of protein in all my lanes. In my control samples (no hypoxia), my housekeeping antibody (alpha tubulin) levels are pretty similar over the 24 hour period, reassuring me that I do have similar levels of proteins in my lanes. However, in my hypoxic samples, this is not the ca…

Is it possible to ligate a vector cut by 2 different blunt ended restriction enzymes?

Detailed answers would be appreciated: 1) What is the difference, in terms of data obtained, between Coomassie Blue staining and Western blotting. 2) What kind of important information can be obtained from purifying a protein from the native species itself? 3) Why is it not a good idea to use too much of template DNA for a PCR reaction? Any help would be appreciated, thank you

Hi all, long-time reader, first-time caller. I'm doing a multi-step purification to purify a protein (or proteins!) responsible for a particular biochemical activity. I do not know the identity of the protein, so I am doing the purification by following the activity only. Here is my purification table. _____Step_____________________Total protein_____Specific activity ____Yield____Fold_________ __________________________________(mg)____________(units/mg)__________(%)___purification___ Processed crude fraction___________57.6______________35_______________100_______=1_____ Ammonium sulfate___________________36.9______________74_______________87________2.13…

Couldn't we try to collect cow poop in a major tank for every area and leverage the methane? Furthermore we could collect all biological waste in another big tank and if we add "thermotoga neapolitana" bacteria in that tank, we can get hydrogen out of it. If we combine oxygen, hydrogen and methane, we can produce methanol. The profits generated from the methanol can be used to invest in this tanks and poop infrastructures that would be needed in the first place. Why methanol ? About the methanol economy: http://www.technologyreview.com/biztech/wtr_16466,296,p1.html Apart from that, we really need a new poop infrastructure anyway for at least 2.6 billion pe…

hi. do you know if platelets respond to mtt assay? what other techniques or experiments can i use to determine viability of platelets? will platelets last 3 days in a culture plate given standard media and conditions? also, how do you count platelets? can i use a hemacytometer? thank you!

Whilst I am not trying to over simplify the multiplicity of causes that can contribute to coronary heart disease, I wonder if prevention is better than a cure. For example, if people at risk took regularly high levels of S methyl cysteine that is found in garlic and turmeric, could they avoid the deleterious effects of high blood cholesterol levels and the increased risk of atheroma, aneurysm or thrombosis? Link For example, this article suggests that a synergistic effect of S methyl cysteine and another chemical can cause a measurable effect in reducing cholesterol in patients: Link to Abstract However, how many people even realised that consuming S…

Please resend this to other people , many people together can solve this tragedy, please suport this letter of Vinko Rajic, help to find out what is this about: I am maybe the only really telepathic person in the world. You should not think that I could be Schizophrenic. STOP PSYCHO TERROR ON ME , PSYCHOPATS ARE PSYCHO KILLING ME, I AM TOTALY MAD OF PSYCHO TORTURE. Can you please resend this email to other scientists, please save me of this terror. Can you please give me advice , what I can do against gang that torture me for last 10 Years. They cost me all important things in my life for last 10 years. I can not pay back money to CSN, I am this winter going…

hi, if i have sequences of integers (have meaning in ordering) , and i want to make pairwise alignment between each two sequences. what is the best way to encod these sequenes? is converted each sequence of integers into nucleotides sequences? or convert each sequence of integers into amino acid sequences? or convert each sequence of integers into nucleotides sequence, then convert it into amino acids sequences? is the length of sequences has relation with the method of the converting? many thanks