P2x3 Antibody Used In Flow Cytometry

April 23, 2018

Biology
Virginia Commonwealth University
PhD Student

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Company:

Novus Biologicals

Product Name:

anti-P2X3 Antibody

Catalog Number:

NB100-1658

The general aim of my research is to determine how P2X3 is involved in mast cell activation. This particular experiment involved the knocking down of P2X3 by specific siRNA. An important part of knockdown experiments is to prove that the targeted protein is actually decreased compared to the scramble control. Using this product we saw that P2X3 expression in bone marrow derived mast cells is decreased about 70% in the knockdown cells compared to the scramble siRNA control cells. This product was selected because it was already validated for flow cytometry use.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse bone marrow derived mast cell

Primary Incubation

Dilution: 1:10, 60 mins at 4 degrees Celsius

Blocking Agent

Anti-mouse 2.4g2

Secondary Incubation

Anti-rabbit PE conjugated

Tertiary Incubation

N/A

Detection

Flow cytometry

Results Summary

P2X3 is expressed on approximately 90% of our primary mast cells. Knockdown by siRNA resulted in about 70% decreased protein expression.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Works well with two step intracellular staining

The Bad

1:10 dilution, each tube allows for only 4-5 experiments

The Bottom Line

Even though it gets expensive due to the low dilution needs, I have tried other antibodies against P2X3 and this works the best for intracellular staining by flow cytometry.

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