H3K9un (7D7) Monoclonal Antibody

$Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K9un (7D7) Monoclonal Antibody (bsm-53024M). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.$

Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K9un (7D7) Monoclonal Antibody (bsm-53024M). The antibody was 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.

$HeLa cells were stained with H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9un antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.$

HeLa cells were stained with H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9un 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.

$ChIP assays were performed using HeLa cells, H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million cells, using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the coding regions of the MYOD1 and MB genes and for a region 1 kb upstream of the GAPDH promoter, used as positive controls, and for the ZNF510 coding region, the EIF4A2 promoter and the Sat2 satellite repeat region, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using HeLa cells, H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million cells, using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the coding regions of the MYOD1 and MB genes and for a region 1 kb upstream of the GAPDH promoter, used as positive controls, and for the ZNF510 coding region, the EIF4A2 promoter and the Sat2 satellite repeat region, 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
IHC-P
IF(ICC)

Reactivity

Human

Overview
Catalog #	bsm-53024M
Product Name	H3K9un (7D7) Monoclonal Antibody
Applications	WB, IHC-P, IF(ICC)
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Mouse
Source	Monoclonal antibody raised in mouse against a region of histone H3 containing the unmodified lysine 9 (H3K9un), using a KLH-conjugated synthetic peptide.
Clonality	Monoclonal
Clone #	7D7
Isotype	IgG1
Concentration	2ug/ul
Purification	Protein A purified monoclonal 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	Histones are present in the chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called histone code. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases.
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IF(ICC)	1:50-200