For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.
Application
Dilution
Western Blotting
1:1000
IHC Leica Bond
1:200 - 1:800
Immunohistochemistry (Paraffin)
1:100 - 1:400
Immunofluorescence (Immunocytochemistry)
1:800 - 1:3200
Flow Cytometry (Fixed/Permeabilized)
1:100 - 1:400
Chromatin IP
1:50
Chromatin IP-seq
1:50
CUT&RUN
1:50
CUT&Tag
1:50
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #35861.
Specificity / Sensitivity
Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit Monoclonal Antibody detects endogenous levels of histone H3 only when tri-methylated on Lys27. This antibody does not cross-react with non-methylated, mono-methylated or di-methylated Lys27. In addition, this antibody does not cross-react with mono-methylated, di-methylated or tri-methylated histone H3 at Lys4, Lys9, Lys36 or Histone H4 at Lys20.
Species Reactivity:
Human, Mouse, Rat, Monkey
Species predicted to react based on 100% sequence homology:
Xenopus, Zebrafish
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys27 is tri-methylated.
