H3R17me2 Polyclonal Antibody

$ChIP assays were performed using human osteosarcoma (U2OS) cells, the antibody against H3R17me2 (bs-53054R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells per ChIP reaction. A titration of the antibody consisting of 2, 5, 10 and 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. The figures show the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Left Figure: QPCR performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. Right Figure: QPCR performed with primers for the promoter of the active ALDOA gene and for the coding region of the inactive MYOD gene.$

ChIP assays were performed using human osteosarcoma (U2OS) cells, the antibody against H3R17me2 (bs-53054R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells per ChIP reaction. A titration of the antibody consisting of 2, 5, 10 and 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. The figures show the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Left Figure: QPCR performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. Right Figure: QPCR performed with primers for the promoter of the active ALDOA gene and for the coding region of the inactive MYOD gene.

To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3R17me2 (bs-53054R). 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 1:40,000.

A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3R17me2 (bs-53054R) with peptides containing other modifications of histone H3 and H4 and unmodified sequences from histone H3. 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. The figure shows a high specificity of the antibody for the modification of interest.

$Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the antibody against H3R17me2 (bs-53054R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.$

Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the antibody against H3R17me2 (bs-53054R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.

Applications

WB
ELISA
dot-blot

Reactivity

Human

Overview
Catalog #	bs-53054R
Product Name	H3R17me2 Polyclonal Antibody
Applications	WB, ELISA, dot-blot
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against histone H3 containing the asymmetrically dimethylated arginine 17 (H3R17me2(asym)), using a KLH-conjugated synthetic peptide
Clonality	Polyclonal
Isotype	Not Determined
Concentration	Not Determined
Purification	Whole antiserum from rabbit containing 0.05% azide
Storage Buffer	Serum with 0.05% sodium azide.
Storage Condition	Store at -20°C 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
dot-blot