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A synthetic peptide containing RT[Me3-K]QT in which Me3-K corresponds to trimethyl lysine 4 of human histone H3.
Our Abpromise guarantee covers the use of ab71998 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). IP is recommended before WB. This antibody is not ChIP grade. Results shown in the image indicate a slight enrichment at heterchromatic loci. This suggests the antibody may exhibit some cross-reactivity with histone modifications localized at these regions. We would welcome feedback about this antibodies performance in ChIP.|
|IP||Use a concentration of 2 - 5 µg/ml.|
|ELISA||Use a concentration of 0.01 - 0.1 µg/ml.|
ab71998 at 1/100 dilution staining Histone H3 in 3T3 cells by Immunocytochemistry/ Immunofluorescence. A FITC labeled goat anti Rb IgG was used for visualization under fluorescent microscope.
Chromatin was prepared from Jurkat 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 ab71998 (blue), and 20µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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