IGF_1 Detection

[anoopbal]

I tried western for IGF-1 twice, but dint find any bands. THe MW is 17.

I am unsure if there is no IGF_1 in the sample or if I should try to optmize my westerns. Is it possible that if the levels can become too low of a protein that yu might not be able to see it even in a control group? Or you should see band, atleast a light one, for wahetevr protein you look for in your tissue?

Thanks

[moleculardude]

Dear Anoopbal,

Have you tried Immunoprecipitation? That way you could pull down all the IGF-1 and then do western blot on a more concentrated sample.

Also make sure you lyse in the smallest volume possible and load most of it.

Before you do all that however, I would make sure you can see IGF-1 with a control using your antibody.

cheers!