Integrin B1 Staining in HT1080 cells - (Feb/14/2013 )
I am looking at total amount of B1 integrins in HT1080 cells using TIRF microscopy. I used the Huts-4 antibody for active B1 and got good localization at adhesions. However, I have used 2 other antibody to look at the total B1 integrin levels and I see nothing specific... No localization at adhesions... So, if you know of any good antibody for detecting total level of B1 integrin, please share . And if you have encountered similar problem, also please share suggestions/advice. Thanks!
I am fixing with 2% PFA (0.12 M sucrose), and permebealizing with 0.2% triton X
you can check which region they bind to at company websites. they must indicate amino acid numbers. so chose antibodies that bind to where Huts4 binds to. I do understand it is active form of B1, and this region might be hidden inside in inactive form, but still check the binding regions.