I was wondering if anyone used the housekeeping gene actin and faced a problem when performed WB. I have been trying to detect the expression using different sources of Action on purified Human Primary Mast cells but nothing came out.
Bandar Baothman
Action concentration is 200 microgram / ml and I have used different dilutions up to 1/100 diluted with TBS/Milk
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Abbas Raza
check for alpha-tubulin or GAPDH. did you run any other western blots or you are running actin first?
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Abbas Raza
if this is your first time doing westerns first run an SDS gel and stain with comassie to see if you are getting properly lysed protein bands.
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