Contact experts in SDS-PAGE to get answers
1,323 views 2,322 posts
Questions related to SDS-PAGE
Dear all, I am using Dde-Azide Agarose (https://clickchemistrytools.com/product/dde-azide-agarose/) and I am trying to click alkyne-tagged proteins via CuAAC to them (ON on 4C), wash, and elute...
02 February 2020 7,871 2 View
I am doing an SDS PAGE with LAD2 cells lysate but the total protein obtained was 0.4ug/ul when 200ul of RIPA was used to lysed 5 million cells. Sample was lysed for 45 minutes on ice. Kindly help.
02 February 2020 6,604 3 View
Hi there, I'm trying to purify some potentially interesting native secreted protein for a bacteria. I can see some good strong peak on the UV trace following IEX chromatrography using an AKTA...
02 February 2020 4,100 3 View
If I have 3H-labeled samples on SDS-PAGE and stain them with silver stain, will they still be compatible with enhancing and fluorography methods?
31 January 2020 1,059 1 View
Hi everyone... For SDS-PAGE sample preparation,I used 250 microlitres of a seaweed sample with 250 microlitres of treatment buffer. But I didn't get any band/protein fraction in it. So can anybody...
30 January 2020 760 2 View
I ran an SDS-PAGE gel of my total cell samples, each expressed in increasing concentrations of a co-factor to see whether this improves expression of the target protein. I coomassie stained the...
29 January 2020 1,847 3 View
I am performing conjugations where I am adding oligonucleotides to proteins. For my confirmation steps, I am doing SDS-PAGE and staining the gel using Pierce Silver Stain to observe shifts in the...
24 January 2020 9,517 3 View
I made several point mutations on a protein, and found that only one particular Ser to Ala mutation leads to significant change in mobility of protein on SDS-PAGE (WB is attached). The drop in...
17 January 2020 8,927 4 View
17 January 2020 3,742 4 View
Hello, I wanted to quantify some samples but noticed that my protein ladder only became visible halfway through the blot. I found it particularly strange because my sample could transfer onto the...
15 January 2020 431 0 View
I am trying to look at protein in a non-cellular structure using SDS-PAGE. I will be collecting plenty of those structures from the model organism on different days. Is it wise if I collect...
13 January 2020 3,010 6 View
I overexpressed soluble cytochrome c in e.coli. After Ni-NTA. I am getting two protein bands on SDS-PAGE - the one which matches my theoretical protein molecular weight 40 kda is having a fainter...
04 January 2020 1,782 2 View
01 January 2020 5,553 2 View
01 January 2020 7,204 4 View
I am working with WD40 genes in cyanobacteria. I have cloned a WD40 gene of size 800bp in the pET28a vector and expressed it in BL21 (DE3) cells. I have induced it with 0.5 mM of IPTG. The...
01 January 2020 5,573 3 View
01 January 2020 2,755 4 View
Hi, I am facing this problem sometimes with PAGE transfer. After I stained with Poncuaue I saw some white horizontal lines on my membrane which are consistent with my sandwich holes. I can...
31 December 2019 2,299 3 View
1) Hi, Do you know that NIBS 98/574 standard is used in SDS-PAGE? (in this standard COA, it's not mentioned that the above standard is suitable for SDS-PAGE or not?) 2) why my recombinant human...
29 December 2019 2,001 5 View
12 December 2019 8,868 5 View
12 December 2019 2,470 3 View
Hi all, I tried to express membrane protein MntBC using vector (H-mntB, mntC-H) in E.coli BL21 Gold. Grew cells in TB + 40% Glycerol amp 3L to OD 1.75 under +37C. Induct with 0.5mM IPTG - 300C...
12 December 2019 9,544 1 View
In my limited experience in proteomics it seems obvious that trypsin digestion and sample preparation is highly biased towards proteins of certain properties. Despite being the most abundant...
12 December 2019 10,025 3 View
This has really been quite frustrating. Every time I load my samples (20 to 45 ug per well), I see lines form in the resolving gel and they interrupt the run. The proteins just accumulate on that...
11 December 2019 6,957 10 View
I have recently adapted the OPA degree of hydrolysis quantification method from Nielsen 2001. I have gotten absorbance readings for standards and hydrolyzed soy proteins that make sense, but I am...
10 December 2019 5,930 8 View