NBP2-54610

antibody from Novus Biologicals

Targeting: H3C14 H3, H3.2, H3/m, H3F2, H3FM, HIST2H3C, MGC9629

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Protein array (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

NBP2-54610

Provider

Novus Biologicals

Product name

Rabbit Polyclonal Histone H3 Antibody

Antibody type

Polyclonal

Description

Peptide affinity purified.

Reactivity

Human, Mouse, Zebrafish

Host

Rabbit

Isotype

IgG

Vial size

50 ug

Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Western blot

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K4me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.

ELISA

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

ELISA: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] [Trimethyl Lys4] Antibody [NBP2-54610] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K4me3. 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 1:18,350.

Immunohistochemistry

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Immunohistochemistry: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] Antibody [NBP2-54610] - HeLa cells were stained with the antibody against H3K4me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] [Trimethyl Lys4] Antibody [NBP2-54610] - ChIP assays were performed using human HeLa cells, the antibody against H3K4me3, and optimized PCR primer pairs for qPCR. Figure shows the recovery, expressed as a percent of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.

Other assay

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Dot Blot: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] Antibody [NBP2-54610] - One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure shows a high specificity of the antibody for the modification of interest.