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H3K4me1 (34A1) Monoclonal Antibody

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using the H3K4me1 Monoclonal Antibody (bsm-53013M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the molecular weight marker (in kDa) is shown on the left.
Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using the H3K4me1 Monoclonal Antibody (bsm-53013M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the molecular weight marker (in kDa) is shown on the left.
HeLa cells were stained with H3K4me1 (34A1) Monoclonal Antibody (Cat. No. bsm-53013M) 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 labelled with the H3K4me1 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 is shown on the right.
HeLa cells were stained with H3K4me1 (34A1) Monoclonal Antibody (Cat. No. bsm-53013M) 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 labelled with the H3K4me1 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 is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K4me1 (34A1) Monoclonal Antibody (Cat. No. bsm-53013M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the monomethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
To test the specificity an ELISA was performed using a serial dilution of H3K4me1 (34A1) Monoclonal Antibody (Cat. No. bsm-53013M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the monomethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using HeLa cells, H3K4me1 (34A1) Monoclonal Antibody (bsm-53013M) 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 1, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. 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, H3K4me1 (34A1) Monoclonal Antibody (bsm-53013M) 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 1, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Bioss unconjugated antibody vial

Applications

  • WB
  • ELISA
  • IHC-P
  • IF(ICC)

Reactivity

  • Human
Overview
Catalog # bsm-53013M-50ug
Product Name H3K4me1 (34A1) Monoclonal Antibody
Applications
WB ELISA IHC-P IF(ICC)
Reactivity Human
Specifications
Conjugation Unconjugated
Host Mouse
Source Monoclonal antibody raised in mouse against histone H3, monomethylated at lysine 4 (H3K4me1), using a KLH-conjugated synthetic peptide
Clonality Monoclonal
Clone # 34A1
Isotype IgG2b
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
IHC-P 1:200-400
IF(ICC) 1:50-200

Frequently asked questions

Which applications is H3K4me1 (34A1) Monoclonal Antibody (bsm-53013M) validated for, and at what dilution?

bsm-53013M is validated for WB (1:300-5000); ELISA (1:500-1000); IHC-P (1:200-400); IF(ICC) (1:50-200). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does H3K4me1 (34A1) Monoclonal Antibody (bsm-53013M) react with?

Tested reactivity: Human.

How should bsm-53013M be stored?

Store at -20C for 12 months. Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

What immunogen was used to raise H3K4me1 (34A1) Monoclonal Antibody (bsm-53013M)?

Monoclonal antibody raised in mouse against histone H3, monomethylated at lysine 4 (H3K4me1), using a KLH-conjugated synthetic peptide

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