DATASHEET
Host:
Mouse
Target Protein:
H3K27me3
Clonality:
Monoclonal
Isotype:
IgG1
Entrez Gene:
8350
Swiss Prot:
P68431
Source:
Monoclonal antibody raised in mouse against histone H3 trimethylated at lysine 27 (H3K27me3), using a KLH-conjugated synthetic peptide.
Purification:
Protein A purified monoclonal antibody in PBS containing 0.05% azide.
Storage Buffer:
0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage:
Store at -20°C for 12 months.
Background:
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
VALIDATION IMAGES
Histone extracts (15 μg) from HeLa cells were analysed by Western blot using H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) diluted 1:1,000 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.
HeLa cells were stained with H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) and 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 labelled with the H3K27me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse secondary antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M). The wells were coated with peptides containing the unmodified H3K27 region as well as the mono-, di- and trimethylated H3K27 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.