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Questions related to SDS-PAGE
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 6,877 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 1,153 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 4,469 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 3,104 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 1,849 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 3,093 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 5,490 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 3,589 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 9,561 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 1,259 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 2,299 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 3,824 3 View
Hello all, so I'm trying to purify a protein complex with multiple subunits for the very first time, and after eluted with different concentration of imidazole there are multiple bands on my...
27 April 2021 6,637 5 View
What happen that caused these white spots/build up? Just some background if needed, I cast my own gels and I ran two 15% SDS Page gels today. One of them came out perfect while my second gel...
24 April 2021 8,095 3 View
I will try to be as thorough as possible. So it's been over a year or so and I went back to pouring various % SDS Page Gels and I keep running into problems I didn't have before as seen below. The...
20 April 2021 3,160 3 View
Polyacrylamide gel that is a component of SDS PAG, my concern is to figure out the concentration of polyacrylamide gel that can be used for SDS PAGE.
20 April 2021 4,186 3 View
I used to dissolve my sample of collagen in different solutions such as 1M NaOH, 0,1M acetic acid, distilled water befor mixed it with laemmli buffer. And also I tried a different gel...
18 April 2021 7,035 2 View
Target bacterial operon is reported to be co-transcribed and co-translated in wild type host. However, protein expression analysis (SDS PAGE) indicates translation of only first gene in frame...
16 April 2021 3,891 3 View
When I was doing a SDS-PAGE upto 25kD the bands of the protein ladder got resolved properly , below the 25kD the 20, 15 and 10kD bands did not resolved and the tracking dye of the samples was...
13 April 2021 8,815 6 View
Does anyone know if there is any condition or modification of protein that will prevent the protein band to be shown on SDS-PAGE gel after silver staining? I recently produced a human protein...
09 April 2021 6,849 3 View
Hello I use the Bradford method to measure the amount of protein expressed by E.coli BL.21. My problem is that despite the R² = 0.9814, the average absorption read at 6 and 21 hours after adding...
09 April 2021 2,085 4 View
Hi all, I've been working with HL-60 cells for ~2 years in our lab. I've been able to successfully differentiate them to a neutrophil-like phenotype using DMSO (1.3% final concentration, 5 days...
06 April 2021 1,818 3 View
In order to produce a chimeric antibody, I transformed HEK293T cells with two plasmids: one for heavy chain and the other for light chain IgG. The supernatant was purified in a Protein G-Sepharose...
29 March 2021 4,662 8 View
I'm carrying out B-cell isolation from mouse tissue by cell sorting using 2x biotinylated probes. One is a wild-type protein and the other has a single point mutation to reduce binding of B-cells...
26 March 2021 4,415 3 View