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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, tri methylated at K36.
(Peptide available as ab1785.)
For detection of methylated histone H3
Our Abpromise guarantee covers the use of ab9050 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration.|
|CHIPseq||Use at an assay dependent concentration. PubMed: 19581485|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K36) peptide (ab1785).|
|ChIP||Use 4µg for 106 cells.|
|ChIP/Chip||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 0.5 - 10 µg/ml.|
ab9050 staining Histone H3 (tri methyl K36) in human differentiated haematopoietic stem cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/400) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat polyclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.
Gating Strategy: Isotype negative control (white).
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab9050 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH (active) and MYO-D (inactive) promoters and over the ý-Actin gene (active). Schematic diagram of the ý-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
ab9050 staining rat liver tissue sections by IHC-P. The section was formaldehyde fixed and subjected to heat mediated antigen retrieval in pH 6.0 citrate buffer prior to being blocked with 5% serum for 30 minutes at 45°C. The primary antibody was diluted 1/500 and incubated for 45 minutes at 20°C. A HRP conjugated goat anti-rabbit was used as the secondary
ab9050 staining mouse kidney tissue sections by IHC-P. The section was formaldehyde fixed and subjected to heat mediated antigen retrieval in pH 6.0 citrate buffer prior to being blocked with 5% serum for 30 minutes at 20°C. The primary antibody was diluted 1/500 and incubated for 45 minutes at 20°C. A HRP conjugated goat anti-rabbit was used as the secondary
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