Cross reactivity with H2AX from human based on the sequence homology with the immunogen. Reactivity against homologues from other sources is not known.
Purification
H2AX pS139 is directed against the phosphorylated form of human H2AX protein at the pS139 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity with non-phosphorylated human H2AX is minimal by ELISA and western blot.
Endotoxin Level
Low Endotoxin : No
Immunogen
Anti-H2AX pS139 purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a phosphorylated synthetic peptide corresponding to the C-terminal region containing serine 139 of human H2AX protein. Immunogen Type: Peptide
H2AFX
Reactivity: Human
WB, IF, ICC
Host: Rabbit
Polyclonal
unconjugated
Application Notes
Anti-H2AX pS139 antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 15 kDa in size corresponding to phosphorylated H2AX protein by western blotting in the appropriate stimulated tissue or cell lysate or extract. ELISA Dilution: 1:5.000 Western Blot Dilution: 1:1.000
Restrictions
For Research Use only
Format
Liquid
Concentration
1 mg/mL
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Handling Advice
Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
Storage
-20 °C
Storage Comment
Store vial at -20 °C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below.
Expiry Date
Expiration date is one (1) year from date of opening.
Rivera-Casas, Gonzalez-Romero, Garduño, Cheema, Ausio, Eirin-Lopez: "Molecular and Biochemical Methods Useful for the Epigenetic Characterization of Chromatin-Associated Proteins in Bivalve Molluscs." in: Frontiers in physiology, Vol. 8, pp. 490, (2017) (PubMed).
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Background
Histones play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA is wrapped around histone-groups, consisting of the core histones H2A, H2B, H3 and H4. As a reaction on DNA Double-strand breaks (DSB) H2AX becomes phosphorylated on serine 139, called gamma-H2AX. ATM, ATR and PRKDCs, kinases of the PI3-family, are responsible for this phosphorylation. The modification can happen accidentally during replication fork collapse, exogenous genotoxic agents, may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes, in the response to ionizing radiation but also during controlled physiological processes such as V(D)J recombination. Mutagenesis experiments have shown that the modification is necessary for the proper formation of ionizing radiation induced foci in response to double strand breaks, but is not required for the recruitment of proteins to the site of DSBs. Gamma-H2AX is a sensitive target for looking at DSBs in cells. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. The role of the phosphorylated form of the histone in DNA repair is under. Anti-H2AX pS139 is ideal for researched interested in Histones, DNA Damage and Repair, and Epigenetics. Synonyms: H2AFX anitbody, H2A.x, H2AX, histone H2A, gamma-H2AX, H2A histone family member X antibody