H2AW Rabbit Polyclonal Antibody

CAT#: TA347207

Rabbit Polyclonal H2A pan Antibody

Product Images

WB was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2 ) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H2Apan. The antibody was diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

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  Determination of the titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H2Apan in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:32, 500.

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  ChIP assays using HeLa cells (sheared chromatin from 1 million cells). A titration of the antibody consisting of 1, 2, 5, and 10 ug per ChIP experiment was analysed. IgG (5 ug/IP) was used as negative control. qPCR primers were for the GAPDH and EIF4A2 promotersas negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeatas positive controls. Image shows the recovery, expressed as a % of input (the relative amount of IP'd DNA compared to input DNA after qPCR).

Specifications

Product Data

• Applications: ELISA, WB
• Recommended Dilution: ChIP (1-2 µg/ChIP); ELISA (1:100-1:1,000); Western blotting (1:2,000)
• Reactivities: Human
• Host: Rabbit
• Isotype: IgG
• Clonality: Polyclonal
• Immunogen: The immunogen for anti-H2A pan antibody: histone H2A using 2 KLH-conjugated synthetic peptides containing a sequence from the central and the C-terminal part of the protein.
• Concentration: lot specific
• Purification: Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
• Conjugation: Unconjugated
• Storage: Store at -20°C as received.
• Stability: Stable for 12 months from date of receipt.
• Gene Name: histone cluster 3, H2a
• Database Link:
  NP_254280
  Entrez Gene 92815 Human
  UniProt ID Q7L7L0
• Background: Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.
• Synonyms: MGC3165

Reference Data

• Protein Pathways: Systemic lupus erythematosus