Anti-p-Histone H2A.X (Ser139) Recombinant Antibody, clone C6B

Histone H2AX (UniProt: P16104; also known as H2a/x, Histone H2A.X) is encoded by the H2AX (also known as H2AFX) gene (Gene ID: 3014) in human. The histone H2A.X protein is a variant member of the H2A family of histones that is distinguished from other H2A histones by a unique carboxy-terminal sequence. This unique sequence is highly conserved throughout eukaryotic evolution and is rapidly phosphorylated by ATM or ATR at the fourth residue from the carboxy-terminus (serine 139 in mammalian H2A.X) in response to DNA double-strand breaks (DSBs). Histone H2AX plays a crucial role in the maintenance of genome stability and the DNA damage response (DDR). H2AX is ubiquitously expressed in almost all eukaryotic cells and is primarily localized in the cell nucleus, where it plays a crucial role in the packaging and organization of DNA within chromatin. Phosphorylation of H2A.X is important in the formation of a stable repair complex at the site of DNA damage. H2A.X phosphorylation is a very rapid response to DNA damage, occurring within as little as one minute after exposure to ionizing radiation. Phosphorylation of H2A.X occurs irrespective of the cause of the DNA DSBs and phospho-H2A.X has been observed in response to environmental stresses that result in DSBs as well as programmed cellular events, including DNA rearrangement and apoptosis. Histone H2AX phosphorylated at serine 139 (gamma H2AX) is a hallmark of DNA damage, signaling the presence of DNA double-strand breaks and global replication stress in mammalian cells. The small size and great affinity for gamma H2AX, allows for efficient electroporation of this C6B nanobody in living cells and, detection of gamma H2AX foci in non-fixed live cells in real time imaging. (Ref.: Moeglin, E., et al. (2021). Cancers (Basel). 13(13):3317).

Ovalbumin-conjugated linear peptide corresponding to 9 amino acids surrounding phosphoserine 139 from the C-terminal region of human Histone H2A.X.

Quality Control Testing

Evaluated by Immunocytochemistry in MCF7 cells.

Immunocytochemistry Analysis: 2 ng/mL from a representative lot detected phospho-Histone H2AX (Ser139) in MCF7 cells treated with 25 M etoposide for 24 h.

Tested Applications

Western Blotting Analysis: A representative lot detected Histone H2AX in Western Blotting application (Moeglin, E., et al. (2021). Cancers (Basel). 13(13): 3317).

Immunocytochemistry Analysis: A representative lot detected Histone H2AX in Immunocytochemistry application (Moeglin, E., et al. (2021). Cancers (Basel). 13(13): 3317).

ELISA Analysis: A representative lot detected Histone H2AX in ELISA application (Moeglin, E., et al. (2021). Cancers (Basel). 13(13): 3317).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone C6B is an E. coli expressed recombinant monoclonal Alpaca antibody that specifically detects Histone H2A.X phosphorylated on serine 139.

Purified E. coli expressed recombinant monoclonal Alpaca antibody in PBS without preservatives with 10% glycerol.

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.