drockhere

Hi all,

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?

Hi there,
I am fairly new to this field, and I have a question about some conversions.
For my research I am using 100ul of a growth factor (of 5ug/ml concentration) in 5 ml of media.

How do i calculate the amount of micrograms I have in the media?

100ul/5000ul is 0.02 ug or 20 nanograms, but how do I use the concentration of the growth factor too while calculating the micrograms?

Thanks a lot, appreciate all the help!