Hello wise scientific community,
I am trying to measure the concentration of Spike Protein ( whole protein) expressed by my cell line transfected with a pC DNA 3.1. I can see a nice band by Western Blot, but when using the same cell extracts ( RIPA buffer plus PI cocktail) in ELISA there is no way to see protein, no matter how much I try to concentrate them. I am using 500 ng of recombinant S protein as a standard to start... I might think the amount of protein expressed is out of the detection range of my assay ( only picograms?), but before deciding that, I wanted to see if anybody has an input about it. A more sensitive assay, a different and more efficient way to extract the protein from the cells lysates... Has anybody tried a transmembrane extraction reagent such as Tm Per-50?
Thanks a lot for your help!
Laura
the antibody works for WB might not work for ELISA.
It seems the antibody you used that works for WB, indicated this antibody recognizes denatured protein. you could try to denature the sample then run the ELISA. but to find another antibody is better.
Thank you Junjie, it sounds as a good idea. I will try denaturing the protein.
Best,
Laura
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