confuse about which antibody to use as a first antibody - (Jan/10/2006 )
I am using a chicken cells transfected with a FLAG tag. I am going to use a antiFLAG antibody to detect the FLAG. I am confused about do I must use a chicken anti FLAG?if not I checked the website of sigma, they have rabbit, mouse anti FLAG, which one I should use?
Thanks a lot
flag is the epitope which will be recognized by the antibody. hence you can choose a anti flag raised in chicken or mouse or rabbit. Secondary ab will be respectively anti chicken, anti mouse or anti rabbit.
as you're using chicken cells, il will be better to avoid a chicken anti flag, as the secondary ab will fix lot of prots...
then you have mouse or rabbit.
I would suggest to choose the opposite as your loading control : if your loading control is a rabiit anti GAPDH for ex, then choose a mouse anti flag.
fred 33, thank you so much. I am clear now You are such a nice person and do know so much. Which country you are in? how many years you did research?