H2AX pS139 in response to CPT

Pharmacology
Boston University
Senior Post-doc

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Company:

EMD Millipore

Product Name:

Anti-phospho-Histone H2A.X (Ser139)

Catalog Number:

05-636

H2AX is a histone variant that is required for DNA damage response. Phosphorylation of H2AX on S139 can be induced by ionizing radiation and genotoxic stresses. Here I tested the camptothecin (CPT)-induced H2AX pS139 in cancer cells.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

U2OS cells

Primary Incubation

Dilution 1:500 in PBS/0.05% Tween20/3% BSA blocking buffer. Overnight incubation at 4C

Blocking Agent

3% BSA / 0.05% Tween20 in PBS

Secondary Incubation

Alexa488--conjugated anti-mabbit 2nd antibody at dilution of 1:250 and 1 hour of incubation at room temperature

Tertiary Incubation

NA

Detection

Fluorescent microscope

Results Summary

The 05-636 antibody worked very nicely for IF. The phosphorylated H2AX S139 foci were evident in CPT treated cells, but not in undamaged cells.

Additional Notes

None

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Summary

The Good

Super specific and sensitive

The Bad

None

The Bottom Line

People who wants to work on phospho-H2AX S139 should definitely try this antibody.

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