HIST1H2AD Antibody - BSA Free

Western Blot: HIST1H2AD Antibody [NBP3-11870] - Western blot 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 NBP3-11870. 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.

Immunocytochemistry/Immunofluorescence: HIST1H2AD Antibody [NBP3-11870] - HeLa cells were stained with NBP3-11870 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Chromatin Immunoprecipitation: HIST1H2AD Antibody [NBP3-11870] - ChIP assays were performed using human HeLa cells, NBP3-11870 and optimized PCR primer sets for qPCR. ChIP was performed on 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 IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ELISA: HIST1H2AD Antibody [NBP3-11870] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of NBP3-11870 in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:32,500.

• Reactivity: Hu
• Applications: WB, ELISA, ICC/IF, ChIP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 4 mg/ml

HIST1H2AD Antibody - BSA Free Summary

• Immunogen: Polyclonal antibody raised in rabbit against HIST1H2AD using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminal part of the protein.
• Clonality: Polyclonal
• Host: Rabbit
• Gene: H2AC7
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1-2ug/ChIP
  • ELISA 1:100-1:1000
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Western Blot 1:2000

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide and 0.05% ProClin 300
• Concentration: 4 mg/ml
• Purity: Affinity purified

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: H2AC7