I have to run on SDS-page very diluted C. elegans lysates (around 0.05 mg/ml) for western blot. I have precipitated proteins with 3 volumes of cold methanol, but I'm not sure this procedure is very reproducible. Do you have suggestions?
Maybe you could try a TCA-acetone precipitation of proteins.
ultrafilteration is a good option with Amicon
Do you know where I can find antibodies directed towards C. elegans? I'm interested in antibodies against LC3, beclin-1 and p62 homologues.
04 May 2012 4,064 3 View
Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...
03 March 2021 4,275 4 View
Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue
02 March 2021 5,433 5 View
I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...
02 March 2021 9,157 1 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
This is gel electrophoresis result for total RNA samples with good conc. However, I guess there is a problem with the purity of the samples, i can't figure out why it appears like this? And how...
22 February 2021 338 9 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