Analysis Note

ControlAcid extracts of HeLa cells.

Application

Research Sub CategoryHistones

Research CategoryEpigenetics & Nuclear Function

Immunocytochemistry:
This antibody has been reported by an independent laboratory to detect ubiquityl-Histone H2A in cells fixed with either 3% formaldehyde/0.1% Triton X-100 or methanol (Vassilev, A. 1995).

Chromatin Immunoprecipitation:
Reported by an independent laboratory (Wang, H. 2004).

Use Anti-ubiquityl-Histone H2A Antibody, clone E6C5 (Mouse Monoclonal Antibody) has been published and validated for use in ChIP, ICC, WB to detect ubiquityl-Histone H2A also known as H2AUb, Histone H2A (ubiquityl), histone H2A.

Disclaimer

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.

General description

Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. Histones are modified post-translationally by acetylation, phosphorylation, methylation, and ubiquitination and these modifications regulate DNA transcription, repair, recombination, and replication. Ubiquitylation usually targets the substrate for degradation, although histones H2A and H2B are actually stabilized by a single ubiquitin conjugation. Histone ubiquitination has been correlated with DNA repair and transcription, cellular differentiation, cell cycle regulation, spermatogenesis, protein trafficking, and response to stress. Histone H2A is monoubiquitinated at Lys119 by the PRC-1L complex (Polycomb repressive complex 1-like), which includes Ring1, Ring2, Bmi1 and HPH2. Ubiquitinated H2A normally represents about 15% of H2A, but this value can be as high as 50% in active chromatin. Ubiquitinyl histone H2A has also been associated with transcriptional silencing of large chromatin regions and linked to Polycomb silencing so the function of ubiquitinated H2A remains undefined.

Immunogen

AMA (human epithelial amnion) cell residual nuclear pellet portions.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

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

Physical form

Purified mouse monoclonal IgM in buffer containing PBS with 0.05% sodium azide before the addition of glycerol to 30%. Liquid at -20°C.

Ammonium Sulfate Precipitation

Format: Purified

Quality

Routinely evaluated by western blot on H2A in acid extracts from HeLa and 10T1/2 cells.

Western Blot Analysis:
A 1:500-1:2000 dilution of this lot detected ubiquityl-Histone H2A in acid extracts from HeLa and 10T1/2 cells.

Specificity

Other species not tested.

Does not cross-react with army worm.

This antibody recognizes and is specific for monoubiquityl-Histone H2A (Lys119), Mr ~25 kDa.

Storage and Stability

Stable for 1 year at -20°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap.

Target description

25 kDa