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Does not react withCarp
Synthetic peptide derived from residues 1 - 100 of Human Histone H3, tri methylated at K27.
. Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the corresponding unmodified peptide and also against a peptide corresponding to tri methylated K9 of Histone H3.
Our Abpromise guarantee covers the use of ab6147 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 - 5 µg/ml.|
|IHC-P||Use at an assay dependent concentration.|
|ELISA||Use a concentration of 1 µg/ml. PubMed: 17641388|
|WB||Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K27) peptide (ab1782).|
|Flow Cyt||Use 1-2µg for 106 cells.|
ICC/IF image of ab6147 stained mouse 2 cell embryo. Cells were fixed with formaldehyde, permeabilized with 0.5% Triton, and incubated with ab6147 at 1/50 for 2 hours at 37°C. Blocking was performed in 5% FCS serum for 30 minutes at 37°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG used at a 1/250 dilution. PI was used to stain the DNA (Red).
ELISA using ab6147.
The red line indicates binding to the tri methyl K27 peptide (ab1782). Weak binding (brown line) was seen to the di methyl K27 peptide (ab1781). Binding to the following peptides was not seen:
Unmodified K27 (ab2623),
mono methyl K27 (ab1780),
mono methyl K9 (ab1771),
di methyl K9 (ab1772),
tri methyl K9 (ab1773),
mono methyl K4 (ab1340),
di methyl K4 (ab7768),
tri methyl K4 (ab1342).
This indicates the specificity of ab6147 for tri methyl K27 of Histone H3.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"