AIF separation process

Cell Signaling/Transduction Apoptosis & Signaling
1

Hello, I am using an AIF antibody from cell signalling technologies (#4642) by western blot analysis. The main problem which I am finding is separating the precursor protein (67Kda) from the mature form (57Kda).

I have used both 10% as well as 7.5% gel and went down to the protein concentration of 10 micrograms, but still I am having problems getting distinct bands.

Any suggestions would be appreciated. Thanks

2

Sarahcanon

It sounds like you’re dealing with insufficient resolution between the 67 kDa precursor form and the 57 kDa mature form of AIF. Instead of 10% or 7.5% gels, try using an 8% or gradient gel (e.g., 8–12%), which might offer better resolution for proteins in the 50–70 kDa range. You can also use a different buffer system using Tris Tricine that offers better resolution in LMW range.

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