I am looking for a protein marker of microglia activation which can be picked up by Western Blot in rat brain homogenates of a neuroinflammation disease model. Preferably Iba1.
I have tried several antibodies already without much success and would appreciate any recommendation. If you have experience with another marker besides Iba1, this information is also highly welcome.
Microglia activation is often characterized using MHC II or CD68, but both also label activated macrophages. Using these antibodies on brain slices would be a lot better than with western blot.
Although I never used it myself for this purpose, the wako rabbit polycolonal anti Iba1 is supposed to work fine for WB (see fig 3 in Szabo M, Gulya K.Neuroscience. 2013 Jun 25;241:280-95.). Following Alexandre suggestion, you can also use MAC-2 antibodies to characterized microglia activation in fixed brain slices.
also support Anti Iba1 (Wako Cat.#019-19741), even if we only used it for IHC on sections of rat brains
Rabbit anti-Iba 1 Wako, Neuss, Germany (019–19741) 1: 1,000; at least it works very nicely for mouse homogenates in our lab: Baltanás et al., GLIA 61:254–272 (2013)
Sorry, the antibody works for Westenrblott and for immunofluorescence (in mouse). It is an antirabbit made of.
What percent of gel did you use? I did a WB blot in December with this antibody in cortical tissue and saw no bands, none at all. I din't use the same secondary, mine was from Roche and my substrate was from Perkin Elmer.
Hey Michel,
Did you get chance to test the Abcam ab5076 antibody for Iba1 in brain homogenates?
Just curious, how it work out for you.
Mayur
Hello
Anti-Iba1 it's work fine for WB
http://www.abcam.com/iba1-antibody-1022-5-ab15690.html
i used rabbit polyclonal anti IBA1 antibody (10904-1-AP) from proteintech and it works fine...
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