Biochemistry and Molecular Biology

I am sorry if my question should be common sense, I am a first year student. I am trying to understand exactly what a Western Blot test determines. In my book it says that a protein has a "lock and key fit with its protein target" and is found by dipping the membrane in a solution with an antibody. In my mind this makes it sound like the test locates the protein by seeing if it fits with its antibody....it does not sound like the test just locates the antibody. I recall that Western Blot tests are used when testing for Herpes and HIV but every site that I find describing the specific test for these two viruses says the western blot test does not locate the viru…

I'm reading about this thing called "Sonneborn's cytotaxis," which is also referred to Sonneborn cytotaxis. I have no clue what this is. I'm guessing it's someone's version of cytotaxis? An idea on how things go about moving in cells in terms of chemical and physical dynamics? Anyone have more information on this? Term used in this abstract: http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0050149 Title: The Mother Centriole Plays an Instructive Role in Defining Cell Geometry

Hi, I have expressed several recombinant proteins in E.coli. But, I never encountered a strange problem like this before. Currently two of the proteins I'm trying to express in E.coli are running about 4-5 kDa bigger than expected size. If it had happened with only one protein then I would assume that particular protein property makes it run it little higher. At this point of time I'm thinking of changing the protein sample buffer containing higher percent b-ME and SDS. Please help me to overcome this problem. Rbrgouda:confused:

Hi All, I have a question about adjusting the pH of a buffer solution. I believe it is not correct to adjust the pH of any buffer by simply adding a few drops of HCl or NaOH, this would introduce a new ion into the solution if Na or Cl was not present in the original solution. Is this a correct philosophy or am I just being stupid? Thanks all

Why are we conscious of only some of our nervous signals and not of all at the same time? Obviously, we are only conscious of what is passing "now" in our consciousness, yet at the same time, we have countless memories stored elsewhere in our brain of which we are not aware. Yet memories are nervous signals like any else, except that they are channeled back and forth from neuron to neuron instead of taking part in a more complex calculation. I guess this is kind of similar to a computer: it's either currently processing information, or else has the information stored in its memory. But in a computer, the difference between information being processed and stored inform…

Guys I have a problem in understanding a discussion with my fellow class mates. He said that "Fact is, supplementing with AKG won't do squat unless you are using something like a membrane permeable ester theoretically. The metabolic intermediates exported from the beta-cell mitochondria into the cytosol have the insulin-releasing signal." I know he is wrong because I readed some studies which I know go against what he said. But is he really wrong ? I asked my teacher he told me to follow up these two links for help but I still don't get it. http://www.ncbi.nlm.nih.gov/pubmed/19502541 http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCH-4J02…

Arthropods' "blood" contains Hemocyanin, to transport Oxygen. And, "Oxygenation causes a color change between the colorless Cu(I) deoxygenated form and the blue Cu(II) oxygenated form". Likewise, it is well-known, that Oxygenated Hemoglobin is blue. And, it is well-known, that the sky appears blue, b/c atmospheric Oxygen (& Nitrogen) preferentially scatter shorter (bluer) wavelengths of light. QUESTION: Does all this imply, that "Oxygen is just plain blue" in appearance (since it scatters shorter wavelength light) ??

I don’t even know if this is possible but could you grow an insect to a certain age and then target and or destroy spliceosome activity without affecting that organism too heavily? I just wonder if spliceosome activity pertains to the phenotype heavily in terms of evolution. As genes may shift in activity via them on an evolutionary scale I guess, its hard to work the idea is all. Basically I wonder if you could stop locus swarms from forming if the spliceosome was involved in that transformation by stopping it at that level. Basically that the locus is a certain phenotype that has enough of a selective advantage to persist, but that selection was sharp enough t…

How long does it take for the cell convert glucose into energy once the glucose has entered into the cell? Ie, how much time passes between the glucose molecule's endocytosis and the end of the molecule's electron transport chain?

What is the purpose of Rnase A in P1 buffer in mini prep kits? Is there RNA floating about outside the bacteria?

HI... actually I need some advices from the forum members... i plan to do the qPCR for several gene of interest (GOI)... and i plan to do the relative quantification... from my reading, i have to test the primers efficiency before i start the real assay.. to do the primers efficiency, i have to generate the standard curve for each pair of primers.. from my reading, i found out that normally researcher will use the 10-fold serial dilution.. so, my questions is : 1) can i use 2-fold @ 5-fold serial dilution of my template??? because i have tried 10-fold dilution but the results is not so good..i think because of my GOI is not highly expressed because the Cq valu…

There is kind of a catch 22 going on. In order to figure out what a gene is people use databases and find homologies in their sequence of interest. How did it start? For example, how did they go about figuring out that haemoglobin is haemoglobin? So, there's mRNA floating about in our erythrocytes, ok great, but how can someone have the slightest idea that this mRNA codes for a protein that binds oxygen. I just don't get how one can go about figuring out for what a piece of mRNA codes. Let's say I isolate mRNA from some bacterium and then I say ok what now. Without using any databases how do I go about figuring out what this mRNA could be. There's all these p…

Hello all, I am looking for a good DNA Gel Imager for use in genotyping my various mouse colonies, cloning and a few other techniques as they come up. Basically all I need is a UV light box and a camera, but I have noticed that there is a wide array of imagers available. The top of the line seems to be Alpha Innotech's Red - but at $10k it's a little out of my price range. I am looking to spend about half that but I am curious as to what people are using in their labs right now. What I need: High rez camera Light source capable of exciting EtBr-stained DNA bands in agarose and polyacrylamide gels. Reasonably small footprint Easily networked or attached/at…

Why is there a change in pH after freeze drying Goat IgG with ammonium bicarbonate buffer but no pH change for rabbit IgG when freeze dried the same? Thanks

Homework question i need help with please CD Spectroscopy is a powerful tool for studying protein secondary structure. Provide detailed narratives/figures to explain the theory, instrumentation, protocol, and anticipated data for structure determinations when (1) PolyGlu, (2) PolyVal, & (3) PolyGly are studied. I understand the theory and instrumentation behind CD, but not the last part about anticipated data for structure determinations of the three listed proteins. Any clarity would be greatly appreciated.

While I'm still interested in obtaining a response to my other post (http://www.scienceforums.net/forum/showthread.php?t=46432), I would now like to ask something more specific regarding bacterial metabolism. How do bacteria go from keto acids to amino acids? Thank you! Merged post follows: Consecutive posts mergedMy bad! I think it's actually the other way around: amino acids are converted into keto acids. Source: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2293160/pdf/2625-07.pdf, FIG. 1.

Hi everyone! I am a FR1 Biochemistry student in Michigan and this is my first post on this forum. I hope you guys will be able to help me figure out some metabolic pathways in E. coli. I am writing a paper on the production of long-chain alcohols by E. coli. The main reference that I'm using is a paper by Zhang, Liao and others of UCLA (http://www.pnas.org/content/105/52/20653.full.pdf+html?sid=1d98aadd-4d83-402b-aa5c-f0bf12e0ff53). These researchers have modified E. coli's lac operon and have added KIVD and ADH6 genes, as well as overexpressing genes already present in the bacterium. I know there are a lot of different pathways that interact to make the alcoh…

i have no background in science really, but have 1 module of it in uni, i need to talk about what enzymes are, include about temperature, equilibrium, non-protein (conjugated proteins) and protein ones (globular proteins), their structure (tertiary which is three dimensional). say what they are used for and why the body needs them. and i genuinel dont have a clue can then talk about how substrate can alter the rate of reaction. anyone help??

How would you detemine the length of an amino acid chain from a formalin titration of a hydrolyzed an unhydrolyzed peptide. My ratio of carboxylic groups came out to be 2.5, but how do i determine the chanin length givien this information?

Hi all! I'm having trouble visualizing a question that asks to compare double-stranded DNA in a solution of PH 7 with a low salt concentration or in water with no ions. In which solution would DNA have a lower melting temperature? I know that DNA is a polar molecule that is negatively charged. Would the ions in the first solution neutralize the DNA and allow it to more easily dissociate (lower melting temperature) than the DNA in the water that wouldn't neutralize the molecule? Or does it have to do more with which one allows it to more easily hydrogen bond?

Hi! I have read many books and gone through few video tutorials but I am still unable to figure it out why actually lagging strand is formed? Thanks

hi, I can really use some help. How would you convert ethanol substrate from 50 microlitres to micromoles/ unit? i dont know if this will help but the ethanol substrate was made as follows 1 mL of ethanol (95%v/v) was diluted with 199 mL of distilled water making the final concentration of the stock 100mM. 15 mL of this stock was thenplaced with 85mL of a buffer solution to make the final concentration 15mM. This stock was used for the enzyme assays.

I didn't know where to post this Thread but the main point is: Do animals get cancer in their natural habitats in fewer numbers than humans? If not, what are the factors that cause the decreased rate of cancer?

Hi does anyone know where to access really good info on the differences between goat and rabbit IgG? Thanks

Why does goat IgG increase its pH after freeze drying in ammonium bicarbonate but rabbit IgG does not?