Monoclonal antibody raised in mouse against histone H3 trimethylated at lysine 36 (H3K36me3), using a KLH–conjugated synthetic peptide

Cross reactivity of the monoclonal antibody directed against H3K36me3
To test the specificity an ELISA was performed using a serial dilution of the monoclonal antibody against H3K36me3 (Cat. No. 25249). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure 1 shows a high specificity of the antibody for the peptide containing the modification of interest.

Immunofluorescence using the monoclonal antibody directed against H3K36me3
HeLa cells were stained with the antibody against H3K36me3 (Cat. No. 25249) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 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.

50 µg/50 µl

PBS containing 0.05% azide and 0.05% ProClin 300

Human

Does not cross–react with H3K9me3, H3K36un, or H3K36me1. Some cross–reactivity with H3K36me2 at high concentrations.

Protein A/G purified

synthetic peptide

Aqueous buffer solution

Store at –80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at –20°C for at least one month.

ChIP (1 – 5 µg/ChIP)
ELISA (1:3000)
DB (1:10,000)
WB (1:1000 – 1:2000)
IF (1:500)

The optimal antibody amount per IP should be determined by the end–user. We recommend testing 1–5 µg per IP

Avoid freeze/thaw cycles

Antibodies

Antibody