H3K36me1 Polyclonal Antibody

$U2OS cells were stained with the Bioss antibody against H3K36me1 (cat. No. bs-53131R) and with DAPI. Cells were fixed with 4% formaldehyde for 20\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum. Figure A: cells were immunofluorescently labeled with the H3K36me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure B, C, D, and E: staining of the cells with the H3K36me1 antibody after incubation of the antibody with 2 ng\/\u03bcl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K36, respectively.$

U2OS cells were stained with the Bioss antibody against H3K36me1 (cat. No. bs-53131R) and with DAPI. Cells were fixed with 4% formaldehyde for 20\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum. Figure A: cells were immunofluorescently labeled with the H3K36me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure B, C, D, and E: staining of the cells with the H3K36me1 antibody after incubation of the antibody with 2 ng\/\u03bcl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K36, respectively.

$Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K36me1 (cat. No. bs-53131R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.$

Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K36me1 (cat. No. bs-53131R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K36me1 (cat. No. bs-53131R) with peptides containing other modifications of histone H3 or the unmodified H3K36. 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.

To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K36me1 (cat. No. bs-53131R), 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 1:46,000.

$ChIP assays were performed using human U2OS cells, H3K36me1 Polyclonal Antibody (bs-53131R) and optimized PCR primer pairs for qPCR. ChIP was performed with a kit, using sheared chromatin from 1.5 million cells and stringent washing conditions. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoter of the constitutively expressed GAPDH gene and for the inactive gene MYOD. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using human U2OS cells, H3K36me1 Polyclonal Antibody (bs-53131R) and optimized PCR primer pairs for qPCR. ChIP was performed with a kit, using sheared chromatin from 1.5 million cells and stringent washing conditions. A titration consisting of 1, 2, 5 and 10 \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 for the promoter of the constitutively expressed GAPDH gene and for the inactive gene MYOD. Figure 1 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
dot-blot

Reactivity

Human

Overview
Catalog #	bs-53131R
Product Name	H3K36me1 Polyclonal Antibody
Applications	WB, ELISA, IF, dot-blot
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 36 (H3K36me1), using a KLH-conjugated synthetic peptide
Clonality	Polyclonal
Isotype	Not Determined
Concentration	0.95ug/ul
Purification	Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
Storage Buffer	PBS with 0.05% sodium azide and 0.05% Proclin300.
Storage Condition	Store at -20°C for 12 months.
Target
Gene ID	8350
Swiss Prot	P68431
Background	A 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
dot-blot