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Questions related to Biochemistry
Hello, I fit a classical dose response curve Y=Bottom + (Top-Bottom)/(1+10^((LogEC-X)*HillSlope)) to my data (not related to inhibitors etc). The model I use is purely qualitative but it fits...
24 February 2021 8,565 5 View
Am in Peptide / Protein Research, may i know what useful analytical information we are getting from 13C & 1H NMR spectroscopy analysis? Somebody getting NMR for smaller proteins, any useful...
06 January 2021 1,365 7 View
Sometimes my western blots have very uniform/even background, other times they're very uneven and patchy like in the attached image. Why is this? I thought my blots were drying out during...
06 December 2020 8,877 5 View
I am currently trying to run solubilised mitochondria extracts from mammalian cells through a gel filtration column in order to separate proteins by size & visualise protein complexes. My...
30 November 2020 1,634 5 View
In the past, I've always used Licor's old blocking buffer that included sodium azide. I would dilute my antibodies in the buffer, stain, re-collect the dilution, and re-use it a few times. The...
29 November 2020 5,967 3 View
I want to reduce the disulfide bridge in my 7kDa protein. My protein has only 1 cysteine at the surface and thus makes a homodimer using a disulfide bridge. I want to reduce the disulfide bridge...
25 November 2020 3,339 5 View
I prepared a 5X of M9 minimal salt and 1000X elemental trace and mixed these 2 solutions to produce carbon-free minimal salt media. Biphenyl was used as a carbon source but was prepped separately...
21 November 2020 6,022 5 View
I'm setting up a liquid handling robot and need a simple assay to test it. Ideally I would like to mix 3-4 reagents and have a absorbance/fluorescence readout on a plate reader. It's also possible...
12 November 2020 7,750 3 View
For western blots, I normally stain with 10 mL buffer with 1:1,000 dilution of my specific antibody. Recently, there is an equipment change in my lab and 10 mL isn't enough volume to cover the...
28 October 2020 2,902 7 View
How difficult is Lysosensor to use in a basic cell culture wet lab? Does anybody happen to have any advice and/or experience with this or similar probes? Thanks in advance!
16 September 2020 8,842 7 View
I must concentrate and purify a lot of dirty lysate and secreted protein samples. I just want to run them on protein gels for western blots. I'm trying to decide the best method of...
11 September 2020 3,052 8 View
I have been searching the literature about this and I have not found an answer. If the atom that accepts a hydride is made more electrophilic, can this speed up the rate of hydride transfer?
30 July 2020 10,048 1 View
I'm trying to perform a BCA assay on some clarified lysate, but the samples appear to become brown, not purple, and the OD562 absorbance is very high. What could cause this? The brown is very...
11 July 2020 7,105 3 View
Various colorimetric assays we use are blue/purple/pink/red but not in green shades. Is there any particular reason for this?
31 May 2020 6,017 1 View
Hello everyone! I have designed a few primers and then a tried optimized the conditions using reference RNA, diferente primer concentrations e annealing temperatures and then calculating the...
26 May 2020 5,838 15 View
Does anyone know if there's a company willing to do all of the following: - Synthesize a custom gene and clone it into a plasmid - Transform E. coli with said gene and develop a protein...
20 May 2020 2,060 9 View
Here is the germane citation: Calvin J Gordon, Egor P Tchesnokov, Emma Woolner, Jason K Perry, Joy Y. Feng, Danielle P Porter, Matthias Gotte. Remdesivir is a direct-acting antiviral that...
15 April 2020 3,764 17 View
In general biophysical experiments are carried out maintaining pH of around 7. For this purpose one can use various buffer systems like phosphate buffer, citrate buffer, tris buffer, bicarbonate...
24 March 2020 3,679 3 View
Hello, I would like inquire on the process of extraction of protein contents from the green pea by means of decantation. What I know is that a decantation is a separation process in which...
23 March 2020 688 4 View
I need to produce formazan to make a standard curve, (as in this paper:Quantitative Determination of Superoxide in Plant Leaves Usi... ) The reaction requires riboflavin and methionine,...
06 March 2020 6,512 3 View
ATP energy-storing molecule as is no doubt an efficient! as not only it is used as energy storage and provider in all forms of life and has probably been around for billions of years. Even in the...
09 February 2020 9,150 6 View
Sometimes I need to centrifuge reactions containing enzymes, however, I've realized I have absolutely no idea if spinning very fast could damage the enzymes. Does anyone have any data on safe...
02 February 2020 7,373 5 View
Hello everybody, I am trying to calculate the irreducible representation of point group 3m (C3v) for NBT (having R3c space group - 161). Could you please give some clarification in this regard?
01 January 2020 6,017 2 View
looking for some best ref. books on Viva in Clinical Biochemistry / Biochemical Analysis / Biochemistry PDF ??
12 December 2019 4,597 4 View