H3K36me3 (11D1) Monoclonal Antibody

$HeLa cells were stained with H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and DAPI. Cells were fixed with methanol and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.$

HeLa cells were stained with H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and DAPI. Cells were fixed with methanol and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 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 of H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M). The wells were coated with peptides containing unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.

$ChIP assays were performed using HeLa cells, H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using HeLa cells, H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Applications

WB
ELISA
IF

Reactivity

Human

Overview
Catalog #	bsm-53021M
Product Name	H3K36me3 (11D1) Monoclonal Antibody
Applications	WB, ELISA, IF
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Mouse
Source	Monoclonal antibody raised in mouse against histone H3 trimethylated at lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide
Clonality	Monoclonal
Clone #	#REF!
Isotype	IgG1
Concentration	1ug/ul
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 Condition	Store at -20C for 12 months.
Target
Gene ID	8350
Swiss Prot	P68431
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.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
IF