Histone H3 Rabbit Monoclonal Antibody for Normalization

Histone H3 Rabbit Monoclonal Antibody for Normalization

The Histone H3 primary antibody can be used as an internal loading control for normalization and is particularly effective when detecting target proteins in nuclear extracts.

The expression of Histone H3, or any housekeeping protein (HKP), should be validated to ensure that its expression does not change under experimental conditions.

Once validated, Histone H3 primary antibodies can be used for the detection of Histone H3 when performing multiplex Western blot detection.

Detect Histone H3 Rabbit Monoclonal Antibody with IRDye® Goat anti-Rabbit or IRDye Donkey anti-Rabbit secondary antibodies.

Other options for housekeeping protein normalization

Reactivity and Specificity

Histone H3 antibody is supplied in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and <0.02% sodium azide.

Do not aliquot the antibody.

Properties Histone H3 Rabbit Monoclonal Antibody (P/N 926-42219)
Species Cross-Reactivity Human, mouse, rat, monkey
Target Molecular Weight 17 kDa
Isotype Rabbit IgG
Specificity/Sensitivity Detects endogenous levels of total Histone H3 protein (including isoforms H3.1, H3.2, and H3.3) and Histone H3 variant CENP-A. Does not cross-react with other core histones. May cross-react with bovine, chicken, D. melanogaster, hamster, xenopus, and zebrafish.
Immunogen A synthetic peptide that corresponds to the carboxy terminus of the human histone H3 protein
Tested Applications Western blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Flow Cytometry (F)

Detection of Histone H3 Rabbit Monoclonal Antibody in HeLa, NIH/3T3, and Cos7 Lysates

Histone H3 Rabbit Monoclonal Antibody detected in HeLa, NIH/3T3, and COS-7 Lysates
Histone H3 Rabbit Monoclonal Antibody detected in HeLa, NIH/3T3, and COS-7 Lysates. Lysates were diluted from 2.5 μg to 156 ng. Lysates were separated on 4-12% Bis-Tris gels, electrophoresed at 200V for 45 minutes in MES Running Buffer, and transferred to nitrocellulose membranes in Tris-Glycine buffer at 100V for 65 minutes. Blots were blocked with Intercept® (PBS) Protein-Free Blocking Buffer (P/N 927-90001), probed with Histone H3 Rabbit Monoclonal Antibody, and detected on an Odyssey® CLx Imager.

Selected P/N: 926-42219

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When using an HKP as your normalization strategy, it’s important to validate the HKP for each experiment to ensure its expression is stable.

Many factors can influence expression including tissue, treatment, and cell density.

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