DATASHEET
Host:
Mouse
Target Protein:
H3K9me2
Clonality:
Monoclonal
Isotype:
IgG1
Entrez Gene:
8350
Swiss Prot:
P68431
Source:
Monoclonal antibody raised in mouse against histone H3, dimethylated at lysine 9 (H3K9me2), 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 H3K9me2 (9B3) Monoclonal Antibody (Cat. No. bsm-53016M) 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 H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M). 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 H3K9me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse 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 H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M). The wells were coated with peptides containing the unmodified H3K9 as well as the mono-, di- and trimethylated H3K9 and the dimethylated H3K4 and H3K27. Figure shows a high specificity of the antibody for the modification of interest.