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Questions related to SDS-PAGE
For my 15kDa protein, what is better the 4-12% Mini Protean TGX gel from Biorad or 4-12% NuPage precast Bis-Tris gel? Is there a significant difference between the two? I'm doing the WB, could the...
29 June 2021 1,773 1 View
hello everyone i have a high background problem in SDS PAGE. i used 2-mercaptoethanol and SDS to denature the IgG, BSA sample (in 1x PBS) and i boiled it for 5 min i run the gel 100 V for 10min...
28 June 2021 1,244 5 View
Dear All, I used (50% Lithium Tetraborate , 50% Lithium Metaborate FLUX and 20% Ammonium Iodide for fusion of PHOSPHATE and CALCIUM samples and FUSION furnaces, Drying oven is fully calibrated,...
23 June 2021 9,876 2 View
Hi! I have purchased a CD34-FITC (IgG) antibody from a company and in order to conjugate it with a gold nanoparticle, I had to purify it and remove all the stabilizing proteins. I used a BSA...
21 June 2021 7,702 6 View
I am trying to my crude enzymes protein sample in SDS PAGE but protein marker (3.5 to 205 kda) band was came while my crude enzymes and BSA 1000 ppm sample was not came the band. Where, I am using...
19 June 2021 2,369 2 View
There are two bands along the length of these gels that mimic the shape of the wells. They show up even in lanes with no sample loaded (the lanes next to the ladder on the gel with two ladders)....
18 June 2021 7,434 3 View
I have data from LC/MS-MS, that gave 55 results, including some contaminations. The main objetive is to try to identify proteins with 56-60 kDa. What are the coverage, #peptides, # PSMs, # Unique...
18 June 2021 6,107 3 View
Given sigma mean and allowable error Is there a relevant formula derivation to give a clear answer
17 June 2021 7,574 2 View
I am using cytiva microspin G25 columns to get rid of excess unicorporated ATP from my protein sample. The protein is 28kDa. The issue that I am facing is that after following the Cytiva protocol...
15 June 2021 2,758 4 View
I was wondering whether CD8 T cells that are present in the spleen are also present in the para-aortic lymph nodes during atherosclerosis. I can not find any article that studies the specific...
14 June 2021 5,047 2 View
Upon imaging Acanthamoeba castellanii cells on a Leica SP8 confocal microscope, I get a moderate to strong level of signal from the nucleolus when trying to image a FISH probe with Atto550 as the...
08 June 2021 6,891 0 View
In a research project I need to confirm the genetic identity of the thrips Frankliniella occidentalis. I would appreciate the help of a fine connoisseur of Tripidae for this task. This help will...
08 June 2021 9,037 3 View
I'm making timelapse recordings on a confocal microscope, but I'm having some difficulty deciding how to normalize the data. For each culture, I recorded a full timelapse with no interference on...
01 June 2021 8,809 0 View
It's just a curiosity that could also help me in my troubleshooting: does Laemmli buffer behave "normally" without the lysate? Like, what happens if I load in the well 20 μL of Laemmli instead of...
27 May 2021 1,167 2 View
I have to perform Western Blot on FFPE tissuse First I did deparaffination, then I added the lysis buffer (20 mM Tris-HCl pH 8, 2% SDS) and I incubated for 20 minutes at 99°C and for 2h at...
27 May 2021 4,486 6 View
Dear RG-community We wonder where we can buy commercial preparative gels for SDS-PAGE. This is a gel with one large slot for the sample and one or two small slots for kD-marker. We do use the...
25 May 2021 3,122 2 View
We have performed SDS PAGE for separating the extracellular protein of staphylococcus aureus resistant strain MRSA... we have followed the aluminium sulphate precipitation protocol for salting out...
18 May 2021 1,861 2 View
Why do all proteins move towards the positive electrode in SDS-PAGE? why do proteins in SDS-PAGE separate according to molecular mass? how can SDS-PAGE be tuned to different optimal ranges of...
17 May 2021 3,103 3 View
I have PEG conjugated antibody in pH 10 borate buffer. I have to run SDS PAGE to determine MW. In this case prior running the gel do I need to do buffer change of the sample? Like borate buffer to...
17 May 2021 5,502 2 View
The composition of the two test samples are given on the day of the exercise. Identify as many proteins as possible on the two gels. Does the number of bands correspond to the number of proteins...
17 May 2021 3,599 1 View
I am trying to purify the galactosaminyl transferrase, LgtD. I used BL21 E coli strain, lysis buffer (EDTA+NaCl+Tris Hcl) and washing buffer (imidazole conc 5 mM) and Elution buffer (20mM...
13 May 2021 9,571 2 View
My SDS-PAGE gel had many blue spots in the region below 40kDa, obscuring all protein bands in this region (see picture). Could someone explain to me what was the problem and how to fix it? The...
09 May 2021 1,270 2 View
I am doing SDS-PAGE using an 8% resolving gel and 5% stacking gel with a Tris-glycine running buffer. During electrophoresis, I get a smeared/tailing dye front which is totally opposite with the...
07 May 2021 2,312 3 View
Hey there, im planning to do an extraction of collagen from decellularized and defatted animal tissue. I would like to extract the collagen by using acidic or enzymatic conditions to obtain telo-...
04 May 2021 3,835 3 View