Anti-phospho-Histone H2A.X, gamma, (Ser139) (H2A Histone Family Member X, H2A.FX, H2A/X, H2AF

Histone H2AX is a 14kD ubiquitous member of the H2A histone family. It contains an evolutionary conserved SQ motif at the C-terminus in eukaryotes. Serine 139 within this motif becomes rapidly phosphorylated to yield a form known as gamma-H2AX in response to double-strand DNA damage. The response of eukaryotic cells to double-strand breaks in genomic DNA includes the sequestration of many factors into nuclear foci. H2AX becomes extensively phosphorylated within 1-3 minutes of DNA damage and forms foci at break sites. Applications: Suitable for use in Immunofluorescence, Immunohistochemistry, Flow Cytometry and Western Blot. Not recommended for use in Immunoprecipitation. Other applications not tested. Recommended Dilutions: Immunofluorescence (IC): 1:500-1:5000, Immunohistochemistry (frozen): 1:1000-1:5000. , Immunohistochemistry (Paraffin): 1:1000-1:5000. Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections. Formaldehyde fixation followed by permeabilization with Triton-X 100., Flow Cytometry: 5ug/1e6 cells in a 150ul reaction, Western Blot: 1:2000-1:10,000, Optimal dilutions to be determined by the researcher. Positive Control: Human HEK293 nuclear extracts, breast carcinoma, HeLa cells, ovarian carcinoma, testicular seminoma. Storage and Stability: May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.