H3K4me1 Polyclonal Antibody

$ChIP assays were performed using U2OS cells, the antibody against H3K4me1 (bs-53037R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 or 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the promoter of the ALDOA gene and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive under normal conditions. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using U2OS cells, the antibody against H3K4me1 (bs-53037R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 or 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the promoter of the ALDOA gene and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive under normal conditions. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3K4me1 (bs-53037R). 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 1:3,800.

$Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using H3K4me1 Polyclonal Antibody (bs-53037R) diluted 1:750 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.$

Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using H3K4me1 Polyclonal Antibody (bs-53037R) diluted 1:750 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 antibody against H3K4me1 (bs-53037R) with peptides containing other modifications or unmodified sequences of histone H3. Other histone modifications include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36 and 79. One hundred to 0.2 pmol of the peptides 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.

Applications

WB
ELISA
dot-blot
ChIP-seq

Reactivity

Human

Overview
Catalog #	bs-53037R
Product Name	H3K4me1 Polyclonal Antibody
Applications	WB, ELISA, dot-blot, ChIP-seq
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 4 (H3K4me1), 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
ChIP-seq