Sounds like a tough problem. A few things might help us to make suggestions.
-What kind of ELISA (direct, indirect, sandwich)?
-What cytokine is of interest?
-Are you using a commercially available antibody, or home-brewed?
-What are you using as a blocking solution?
-Are you putting your antibodies in the blocking solution?
Jason Bubier wrote:
> Hello,
> I am doing an ELISA assay for a cytokine in the serum of mouse. My
> standard curve looks great (4000-63 pg/ml) but it is undetectable in all
> serum samples EVEN when I spike the sample with 250 pg/ml of cytokine
> which works fine in the standard curve! Colleagues have suggested this
> is due to interfering factors from the serum. I have performed dilutions
> of 1:10 and 1:4 but still no detection. Any advice would be greatly
> appreciated as to what parameters I should vary. I was referred to "The
> Immuno Assay" by David Wild which I don't have but am ordering. If you
> have the book let me know what it says about controlling interfering
> substances.
>> Thank You
> Jason Bubier
>> ---
