Blocking Non-specific Binding in Immunoassays

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Agilent

Product Name:

Rabbit Serum (Normal)

Catalog Number:

X090210-8

Rabbit normal serum was the best blocking agent I have used so far to prevent non-specific binding antigens to antibody in my immuno assay reactions. Normal serum provides a sufficient quantity of endogenous proteins to saturate and block nonspecific binding sites on fixed samples or assay substrates, making it useful for immunodetection methods, especially those involving tissue samples such as immunohistochemistry (IHC). Normal sera are also useful as controls for testing general and specific antibody purification methods.

Experimental Design and Results Summary

Application

Immuno assay development especially blocking non-specific binding

Starting Material

Concentrated normal rabbit serum and PBS

Tips

None

Results Summary

I performed following dilution for immunohistochemistry based detection method from paraffin embedded tissue sections; 1. Dilute normal rabbit serum with neutral PBS in the ratio of 1:10.2. Add working solution onto tissue section and incubate at 37°C for 30 minutes.3. Remove the blocking solution, but do not wash the tissue section.4. Follow the protocol for immunohistochemistry based detection using specific antibody according to manufacturers protocol.Overall, assay detection went well by reducing background noise.

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Summary

The Good

Good blocking agent, cost effective.

The Bad

None

The Bottom Line

Highly recommended for immunoassay optimisation.

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