Hi Rasha al-Attar,

I saw your question and Luminex can help you with this. Please send me your contact information at [email protected] and I'll have our Tech Support team and Field Application Scientists help you with your assay.

Charles Martin

Rasha (Q): I'm trying to optimize a multiplex assay but I keep running into a problem.

- I have a protein extract from one sample that has my antigen and protein extract of a similar sample that has everything else BUT my antigen (so a negative control).

Mohammed : So basically in this case you have an antibody (protein extract) and an antigen and lets call this sample 1

Sample 2 contain antibody (protein extract) but does not contain antigen.

Therefore, sample 1 contains an antigen and sample 2 does not contain an antigen.

Rasha (Q): I'm trying to show that the assay is successful in discriminating between the samples containing and lacking the antigen. I was successful in doing it once but I have a problem replicating it.

Mohammed: Therefore, your trying to show that there is a difference between the sample 1 and sample 2

Thus, your trying to determine the difference between the presence of the antigen in sample 1 and the absence of antigen in sample 2.

And your doing this probably because you want to study the effects of that specific antigen on to the antibody (protein extract) Or medically known as the study of the interactions between the antibody and antigen complex.

Rasha (Q): The net FMI of my wild type sample is almost the same as the net FMI of my sample that lacks the antigen.

Mohammed: And your using Fully Auto-Biochemistry Analyzer (FMI-400 )

a medical laboratory instrument designed to measure different chemicals and other characteristics in a number of biological samples quickly, with minimal human assistance.

Rasha (Q): Any suggestions on how I can optimize for the assay?

Mohammed: So my suggestion to you is to read this article and if you still have a problem please contact me.

http://www.statease.com/pubs/optimizeassays.pdf

Rasha (Q): P.S.: my primary antibody is super specific to my antigen. My secondary antibody is commercially bought and can detect ALL the proteins specific to my extract.

I hope this is helpful.

Good luck and you can follow me on facebook also https://www.facebook.com/mohammed.mo.3114 (mohammed mo is the name on facebook)

Salam and take care