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.
Action concentration is 200 microgram / ml and I have used different dilutions up to 1/100 diluted with TBS/Milk
check for alpha-tubulin or GAPDH. did you run any other western blots or you are running actin first?
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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