Hi researchers who study BMP2,
I'm doing western blot for BMP2 recently and really need some help from those who ever done western for this protein!
My major question is what is the real size for BMP2? Or in other words, if you are studing the function of BMP2, for western blot, although there will be propeptide, monomer, dimer... for this protein, which one you are going to detect for your western?
I have this question because when I search for the antibody for BMP2 from different companies, no matter it's poly or monoclonal, the size is very different. Now I have 4 different BMP2 antibody at hand. I tried once to probe BMP2 on the same membrane with two different antibodies. But the band cannot match each other, which means at least one of them is wrong.
I want to hear from people who did western blot for BMP2, what is your band size? How do you confirm that band is BMP2?
Thanks so much!
Shuaichen: You are not alone feeling frustrated by the multiple bands of a small cytokine protein such as BMP2. We've seen similar situation with its family member, TGFb1. This was caused mostly by the carrier glycoprotein (alpha2-macroglobulin or C3 complement) associated with its biosynthetic pathway. and the situation was compounded by the use of traditional WB with a primary Ab and a secondary Ab-HRP which detects trace amount of IgG carried over in the sample (serum). We bypassed this by using direct conjugation of the primary Ab with HRP and the use of non-reducing sample buffer (with or without heating). Please check our recent paper on HK2 cells secreting a spectrum of plasma proteins.
PS: Due to copy right conflict, we cannot post our latest full-length paper on HK2 cells online for free download prior to the publisher's release. However, we can always send the paper to individuals who request directly.
Hi Shuaichen,
I was detecting BMP-2 expression in HeLa lysates and supernatants, and obtained two kinds of bands. For precursor protein I obtained bands with molecular weight around 55-60kDa, and for mature protein 36 kDa. I was working with Santa Cruz Ab: BMP-2 Antibody (N-14): sc-6895 (I found out that this antibody is currently not available - discontinued production).
Hi Agnieszka,
I know the antibody you mentioned. That's another cofusion for me. If you look at BMP2 antibody from different company, the band size differ from 37 to 45 kDa. And you can find publication for these antibody. Some of them use the same cell line but the band they show is diiferent because of the antibody. In such condition, how to confirm which antibody is correct?
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