For optimal ChIP and ChIP-seq results, use 5 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
Simple Western™
1:250
Immunofluorescence (Immunocytochemistry)
1:50 - 1:200
Flow Cytometry (Fixed/Permeabilized)
1:50 - 1:200
Chromatin IP
1:100
Chromatin IP-seq
1:100
CUT&RUN
1:100
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 #87261.
Specificity / Sensitivity
Acetyl-Histone H3 (Lys27) (D5E4) Rabbit Monoclonal Antibody recognizes endogenous levels of histone H3 protein only when acetylated at Lys27. This antibody does not cross react with histone H3 acetylated at Lys9, 14, 18, 23, or 56. This antibody shows some cross-reactivity with acetyl-histone H2B lysine 5.
Species Reactivity:
Human, Mouse, Rat, Monkey
Species predicted to react based on 100% sequence homology:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding acetylated Lys27 of human histone H3 protein.
