Histone H3 [Dimethyl Lys4] Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Histone H3 [Dimethyl Lys4] Antibody [NBP2-59187]

Immunocytochemistry/Immunofluorescence: Histone H3 [Dimethyl Lys4] Antibody [NBP2-59187] - Human osteosarcoma (U2OS) cells were stained with the antibody against H3K4me2 and with DAPI. Cells were fixed with 4% formaldehyde for 20 minutes and blocked with PBS/TX-100 containing 5% normal goat serum. A: cells were immunofluorescently labeled with the H3K4me2 antibody (left) diluted 1:5,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 568 or with DAPI (right), which specifically labels DNA.

Chromatin Immunoprecipitation: Histone H3 [Dimethyl Lys4] Antibody [NBP2-59187] - ChIP was performed with the antibody against H3K4me2 on sheared chromatin from 500,000 HeLaS3 cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody of 0.5, 1, 2 and 5 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. Quantitative PCR was performed with primers for a region upstream of the ACTB and GAS2L1 promoters, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ELISA: Histone H3 [Dimethyl Lys4] Antibody [NBP2-59187]

ELISA: Histone H3 [Dimethyl Lys4] Antibody [NBP2-59187] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3K4me2 in antigen coated wells. 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 1:48,200.

Dot Blot: Histone H3 [Dimethyl Lys4] Antibody [NBP2-59187] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H3K4me2 with peptides containing other modifications of histone H3 and H4 and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.