anti-mouse CD11b in WB - Has anybody used an Ab aganist CD11b that works in WB? (Apr/07/2010 )

Hi everyone,

has anybody used an Ab aganist mouse CD11b that works in WB?

I have the cloned Ab M1/70, and I'm not getting success results in WB, and I would like to know if someone has observed this integrin by this technique with this Ab or any other.

I've been using 8% SDS-page gels in not reducing condition, my work is based in macrophage/monocyte biology so they express CD11b, and the samples have been boiled for 5 minutes at 95ºC.

The transference has been done as usual in methanol buffer.

Thanks a lot for your help, I'll appreciate any piece of advise which you could give me

does the data sheet that came with the antibody say that it is suitable for western blot? it may require native protein (which would be unsuitable for wb).

mdfenko on Apr 7 2010, 05:06 PM said:

Thanks for your help

The Ab is not recommended for WB, but in other cases I've found other Abs which the same recomendation and they have worked.
In this case, the recommendation may be true and the Ab is unsuitble for WB, but I thought about the possibility someone had checked it before than me and obtained good results.

Ok, another question: if I biotinolate the cell surface, would I be able to immunoprecipitate the integrin with this Ab ?
(OK, the data sheet of this M1/70 Ab confirm that it suiteble for immunoprecipitation)

In affirmative answer, could someone give some advice for the protocol?

Thanks in advance