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.
Application
Dilution
Western Blotting
1:1000
Simple Western™
1:10 - 1:50
Immunoprecipitation
1:100
Immunohistochemistry (Paraffin)
1:400 - 1:1600
Immunofluorescence (Frozen)
1:400
Immunofluorescence (Immunocytochemistry)
1:400 - 1:800
Flow Cytometry (Fixed/Permeabilized)
1:400 - 1:1600
Chromatin IP
1:100
Chromatin IP-seq
1:100
CUT&RUN
1:100
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 #69994.
Specificity / Sensitivity
NF-kappaB p65 (D14E12) Rabbit Monoclonal Antibody recognizes endogenous levels of total NF-κB p65/RelA protein. This antibody does not cross react with other NF-κB/Rel family members. Non-specific staining was observed in pancreatic islets by immunohistochemistry.
Species Reactivity:
Human, Mouse, Rat, Hamster, Monkey, Dog
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu498 of human NF-κB p65/RelA protein.
