β-Actin Mouse Monoclonal Antibody for Normalization

beta-Actin Mouse Monoclonal Antibody

Actin is a commonly-occurring structural housekeeping protein and has been used for Western blot normalization for many years.

You need to validate the expression of actin, or any housekeeping protein (HKP), to ensure that its expression does not change under experimental conditions. Otherwise, quantitation cannot accurately be performed.

Once validated, β-actin primary antibodies can be used for detection of β-actin as a normalization antibody when performing two-color detection.

Detect β-Actin Mouse Primary Antibody with IRDye® Goat anti-Mouse or IRDye Donkey anti-Mouse secondary antibodies.

Other options for housekeeping protein normalization

Reactivity and Specificity

β-Actin 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 β-Actin Mouse Monoclonal Antibody (P/N 926-42212)
Species Cross-Reactivity Human, mouse, rat, monkey, hamster
Target Molecular Weight 45 kDa
Isotype Mouse IgG2b
Specificity/Sensitivity Detects endogenous levels of β-actin protein
Immunogen A synthetic peptide (KLH-coupled) that corresponds to the residues near the amino-terminus of human β-actin
Tested Application Western blot

Actin Used for Normalization

Actin used for normalization in
Presence of Smad2/3 in Xenopus embryo lysates. Xenopus embryos were injected with Smad2 RNA and treated with noggin, truncated BMP receptor (tBR), truncated activin receptor (XAR) or BMP4. Lysates were electrophoresed on 12.5% SDS-PAGE gel and proteins were transferred onto PVDF membrane. Membrane was probed for Smad2/3 and β-actin expression using rabbit anti-smad2/3 and mouse anti--actin monoclonal antibody (P/N 926-42212) followed by detection with IRDye 800CW Goat anti-Rabbit IgG (P/N 926-32211) and IRDye 680RD Goat anti-Mouse IgG (P/N 926-68070). Contributed by Kimberly Wong, SUNY Upstate Medical Center, NY, United States.

Selected P/N: 926-42212

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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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