ELISA Problem

Colin North northcol at ____msu.eduRemove_____
Sat Aug 14 09:59:14 EST 2004


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
> 
> ---



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