hello,everyone!
I want to identify that Schwann cells which isolate from postnatal 1-4day Rats is differentiated when culture in differentiation conditions. By western blotting, I got bands of Actin and Oct6 but not MBP. The primary antibody is mouse anti-Actin(1:5000), Goat anti-Oct6(1:2000) and mouse/rat anti-MBP(1:5000) or MBP(1:2500). Should I increase the primary antibody concentration of MBP?
I tried lots of experiment condition，but couldn't get the bands for MBP.
Does anyone do some similar research about Schwann cells？
Waiting for your help.
Do you have a positive control for your Western blot? You could use brain homogenate from an adult rat, it should contain a lot of myelin (including myelin basic protein). If you don't get a signal with the positive control, then something is wrong with your set-up.
Mr. Holger
Thank you for your advice .May I just use oligodendrocytes to do the positive control?
A positive control is the next step. You may also need to vary the concentration of the primary and secondary antibody and try different blocking agents. it is also worth searching the literature and see if there are any other antibodies you could use. If all else fails, you may have to try immunoprecipitation.
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