I'm a grad student looking at multiple proteins via western blot. The very specific (clean and one-band result) antibodies I have used have lower molecular weights than says on the datasheet from the companies. For example, the result I get from one antibody gives me a very specific band around ~38 kDa with whole cell lysates, where as the company expects you to find a band ~45 kDa whole cell lysate (NE). For a different specific antibody I have a ~50 kDa when the expected band should be around ~58.1 kDa.
I'm suspecting it could be a gel concentration problem...I use a Tris-HCL gel 4-20% from BioRad. Is the gel I'm using an issue/gel electropheresis issue or can it be the protein is undergoing post-translational modification/splicing during transcription level?
Thank you for those who can!
Edited by WestSideGrad, 08 July 2010 - 12:09 PM.