I am a newbie in elisa and I need some urgent help.
I have an ELISA kit that does not have any standards so I am assuming it is for relative quantification instead of finding absolute values.
I need to find the extent of phosphorylation of psmad in the lysates or some kind of fold expression with respect to positive controls.
I was wondering can I use an unphosphorylated antibody of psmad as positive control, find its O.D. Then can I normalize the O.D levels of my sample lysates with respect to the O.D of the positive control to get fold change? Does this normalized intensity seem right?
I also have the total protein content of my lysates from a BCA Assay. So if the above is wrong, can I normalize the O.D values to the total protein content? For this do I simply divide the O.D values by the total protein value? (I have in ug/ml)
Is this right?
Edited by drockhere, 17 November 2012 - 10:16 PM.