Histone H3K4me2 Antibody Description: Rabbit polyclonal antibody recognizing the histone H3 monomethylated at lysine 27. Histone H3 is one of the four proteins that are present in the nucleosome, the basic repeating unit subunit of chromatin, consisting of 147 base pairs of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). Methylation of lysine 27 is associated with transcriptional repression by the Polycomb complex of proteins and the formation of facultative heterochromatin.
Histone H3K4me2 Antibody Immunogen:Synthetic peptide derived from human histone H3 monomethylated at lysine 27.
Histone H3K4me2 Antibody Formulation:Affinity-purified IgG (1 mg/ml) in PBS, pH 7.3, with 0.02% sodium azide and 50% glycerol.
Histone H3K4me2 Antibody Quality Control:This antibody has been validated for use in IF, IHC and Western blot. It has also been validated for specificity by dot blot and histone peptide array and it has been confirmed that this antibody:
Is specific for H3K4me2
Has no cross-reactivity to related modifications
Is unaffected by nearby modifications
Histone H3K4me2 Antibody Storage and Stability:Stable for 2 years at -20°C from date of receipt.
Histone H3K4me2 Antibody Application NotesRecommended dilutions:
ChIP 2-5 µg per 1x10^6 cells
IF / IHC 1:50 - 1:200
IP 2-5 µg per IP
WB 1:500 - 1:2,000
Histone H3K4me2 Antibody References:
View technical datasheet for this product. View additional specificity data this product.
SNAP-ChIP qPCR Data: Histone H3K4me2 antibody (1:60 dilution) was tested in a native ChIP experiment with chromatin from HEK-293 cells (~1x10^6 cells). Specificity (left Y-axis) was determined by qPCR for the duplicate DNA barcodes corresponding to each modified nucleosome in the SNAP-ChIP K-MetStat panel (EpiCypher Catalog No. 19-1001, X-axis). Black bar represents antibody efficiency (right Y-axis; log scale) and indicates percentage of the barcoded H3K4me2 nucleosome target immunoprecipitated relative to Input. The antibody exhibited good specificity for the target (<10% cross-reactivity outside H3K4me2) and high efficiency (18.1%). Error bars represent mean ± SEM from two barcoded nucleosome replicates in a single ChIP experiment. (Click image to enlarge)
Specificity Data-Array: EpiTitan Histone Peptide arrays used to analyze the histone modification specificity of Histone H3K4me2 Antibody. Data is shown for the highest reacting singly-modified peptides. For more specificity data, see Additional Peptide Array Specificity Data. (Click image to enlarge)
Specificity Data-Dot Blot: Peptide dot blot used to confirm the specificity of Histone H3K4me2 Antibody. Methylated peptides corresponding to the immunogen and related sites were spotted onto PVDF and probed with the antibody. (Click image to enlarge)
Immunofluorescence Data: HEK293T cells stained with Histone H3K4me2 Antibody (red) and counterstained with DAPI (blue). (Click image to enlarge)
Western Blot Data: HeLa cell extract (HeLa) and recombinant histone H3 (H3 protein) were blotted onto PVDF and probed with Histone H3K4me2 Antibody. (Click image to enlarge)