I had done elisa of bmp2 of elder populations by ELABSCIENCE ELISA KIT by following standard protocol, but every time it shows that OD of serum samples are come lesser than blank means Zero concentration standard.
1st time I thought that Human serum samples are thawing many times that's why proteins are degraded.. but next time i had done this from fresh currently collected sample but again sample problem is happened ..
kindly help me out why this OD values are so low than blank also!
there any reason behind it ..
Monalisa Mahajan , there are many points to check.
In particular, complex samples such as serum often contain various macromolecules such as proteins and lipids. These components may interfere with the binding of antibodies and antigens, leading to signal suppression or even "negative" phenomena (the sample OD is lower than the blank).
From an operational perspective, the adequacy of the plate wash can be enhanced, the time for each wash can be extended, and the wash solution can flow through the bottom of each well to avoid residue. It is recommended to pat the plate dry after the final wash. The color developer and well plate status also need to be checked: the color developer (TMB) should be colorless and transparent; the plate should be used immediately after opening. Prolonged exposure or improper storage may lead to high background.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
how can I overcome interfere with the binding of antibodies and antigens?
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