bs-53118R [Primary Antibody]
H3K9me2 Polyclonal Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: H3K9me2

Clonality: Polyclonal

Isotype: Not Determined

Entrez Gene: 8350

Swiss Prot: P68431

Source: Polyclonal antibody raised in rabbit against the region of histone H3 containing the dimethylated lysine 9 (H3K9me2), using a KLH-conjugated synthetic peptide

Purification: Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.

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.

Size: 50ug

Concentration: 1.755ug/ul

Cross Reactive Species: Human
Mouse
Others
(Arabidopsis, Rice, Tomato)

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Histone extracts (15 μg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) 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.


A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) with peptides containing other modifications of histone H3 and the unmodified H3K9 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.


Mouse NIH3T3 cells were stained with the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) 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 H3K9me2 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.


To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody against H3K9me2 (Cat. No. bs-53118R), crude serum and Flow through. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:103,000.