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Questions related to SDS-PAGE
The power supply is working very well as well as current and still don't separate even the marker
29 August 2021 2,205 3 View
As you can see the bands are separated at the top but they are distorted at the bottom and there is more stain at the bottom while the positive control is at the top. when i satined the gel with...
28 August 2021 9,927 3 View
I am doing purification of a 6xHis-tagged protein and it seems that the protein is not eluted with the traditional 500mM Imidazole in my elution buffer...mostly contaminating proteins are present....
28 August 2021 9,185 9 View
My sample consists of a mixture of proteins. After size exclusion chromatography, the fraction samples were subjected to ESI MS. The SDS-PAGE gel showed the sIze exclusion fraction has many...
20 August 2021 7,189 3 View
Hello everyone We currently facing a problem with our SDS PAGE in our lab, as you can see in the attached file the migration at the end is very chaotic We're using BioRad products for years now,...
19 August 2021 411 4 View
I am trying to lysate BL21 DE3 E. coli for the purification of recombinant proteins Blo t 2 (14.5KDa). I couldnt achieve results in denaturing condition but in native conditions, when using...
13 August 2021 7,785 8 View
I do SDS PAGE at our lab and the results with BSA protein look great. I want to run it with a complex of Polymer and protein. However, everytime I do it, the complex always travels just like the...
11 August 2021 2,073 3 View
Dear colleagues, Please recommend a particular catalogue number of 4-VP for protein electrophoresis. We study storage high molecular proteins in wheat and get different results with different...
10 August 2021 7,874 3 View
After protein extraction i had my sample in to 2 (A&B) tubes and from each tube i made 1 stock and 3 dilutions I got the curve and each dilution's concentration How to measure the final...
06 August 2021 4,644 5 View
Hi, I'm stuck with membrane transferring in Western blot. To analyze the expression level of caspase 3 (cleaved, ~17 kDa), I harvested the cell then lysed by Laemmli sample buffer (6X, w/ DTT)...
27 July 2021 8,335 9 View
Hello, We’ve run a few SDS page gels in the past few weeks but so far, we have not been getting much bands. We bought a PET-21-b+ plasmid and eventually transformed them. After growing it up in...
24 July 2021 7,471 3 View
Dear fellow researchers, I was wondering if different forms and stages of post-translational modifications (such as N-linked glycosylation) on a recombinant protein expressed in insect cells can...
21 July 2021 3,216 13 View
I'm an undergraduate BS-Biology student. Can anyone help me to know a step-by-step method on how to make a tributyrin agar to a tributyrin broth?
18 July 2021 6,242 3 View
I did disassemble/reassemble treatment of HBc VLPs. I collected the disassembled protein fractions using Ion exchange chromatography and reassembled overnight in buffer (NaCl, Hepes, DTT). I made...
15 July 2021 7,829 3 View
I dis disassemble/reassemble treatment of HBc VLPs. I collected the disassembled protein fractions using Ion exchange chromatography and reassembled overnight in buffer (NaCl, Hepes, DTT). I made...
14 July 2021 4,881 2 View
Does anibody know why BL21 DE3 don't express protein? I used pET vector for scFv antibody (only 31 kDa with His-tag) Transformation go well. I grow it with LB and induce with IPTG (new).
13 July 2021 1,333 7 View
I have PEGylated antibody in 6.5 pH phosphate buffer. I want to show change in molecular weight using SDS PAGE. While preparing the sample should I change the buffer of my sample to TRIS-HCl pH...
09 July 2021 1,140 3 View
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 1,765 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 1,231 5 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 7,690 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 2,360 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 7,426 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 6,093 3 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 2,746 4 View