Has anyone ever successfully used mouse serum as the sample in a Western blot? If so, what (if anything) did you use to dilute it?
Thanks so much for your suggestion, Dr. Wicher. I'll give it a shot!
I have used pure serum because the antibodies concentration was very low and had no problem. When i need diluted i use Phosphate buffered saline + BSA 0,1%.
05 June 2016 2,484 11 View
31 December 2014 5,901 2 View
I need to stain rodent implantation sites but then want to use the uteri for histological fixation. I understand that the use of ammonium sulfide renders the tissue unusable for fixation. Any...
05 June 2013 3,023 13 View
I'm looking for a good protocol for spike-in for rat serum miRNA quantification. I've heard that the C.elegans spike-in works well, but don't have an appropriate protocol for it. Any tips would be...
03 April 2013 8,928 10 View
I am running a sandwich ELISA and realised far too late that I have run low on SigmaFast p-npp tablets for my detection step. In the product information insert, Sigma suggests 200 microlitres of...
06 July 2012 1,343 6 View
I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...
25 February 2021 8,404 3 View
I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...
24 February 2021 8,514 3 View
Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...
23 February 2021 8,609 3 View
Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...
23 February 2021 6,671 4 View
I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...
22 February 2021 9,901 3 View
I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...
22 February 2021 6,937 3 View
I ran a western blot using 12% gel and probing for ERK/pERK (cell signaling antibodies). The green is total ERK, the Red is supposed to be phospho-ERK. I see 2 bands below the total ERK that...
22 February 2021 1,777 3 View
Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...
17 February 2021 4,844 4 View
Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...
17 February 2021 1,130 6 View
I accidentally rinsed with methanol after adding my primary antibody to my membrane. I noticed after ~10 seconds and immediately replaced it with PBST (3x15 min washes). Will my antibody signal be...
12 February 2021 5,824 3 View