12 August 2015 7 9K Report

Dear All,

I just wonder that is possible that some protein might not transfer to some type of membrane in Western Blot. for example protein A might transfer to only nitrocellulose, not PVDF.

I have a case that I and my colleague using the same cell line, same protocol, and same antibody to detect different isoform of calcineurin A at 48kda and 60kda. I use PVDF and detect only 60kda, but my colleague use nitrocellulose and detect both 48kda and 60kda.

I'm still thinking what is the issue. The pore size of both membrane are the same 0.45um, so it couldn't be issue. Usually larger molecular weight transfer to membrane more difficultly. But in my case, 60kda has alreaday transfer, but not its lower MW, 48kda doesn't. PVDF binding capcity should allow more protein transfer. Ah I dont know.

Thank you

Cheerio,

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