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Actin Primary Antibodies for Normalization

ß-Actin detected in HeLa, NIH/3Ts, and COS7 cell lysates.

Figure 1. β-Actin detected in HeLa, NIH/3T3, and COS7 cell lysates.

Presence of Smad2/3 in Xenopus embryo lysates.

Figure 2. 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 -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.

β-Actin primary antibodies can be used for detection of β-Actin by Western blot or by In-Cell Western™ assay. These primary antibodies can be used as a normalization antibody when performing two-color detection.

Detection of the β-Actin Rabbit Primary Antibody can be achieved with IRDye® Goat anti-Rabbit or IRDye Donkey anti-Rabbit secondary antibodies.

Detection of the β-Actin Mouse Primary Antibody can be achieved with IRDye Goat anti-Mouse or IRDye Donkey anti-Mouse secondary antibodies.

Note: When lanes are heavily loaded with lysate (>15 µg/lane), a highly expressed internal control protein may not be an appropriate choice for normalization. For quantitative analysis of a low-abundance target protein, a low-abundance internal control is recommended. COX IV is an appropriate choice.

Properties β-Actin Rabbit Monoclonal Antibody (P/N 926-42210) β-Actin Mouse Monoclonal Antibody (P/N 926-42212)
Species Cross-Reactivity Human, mouse, rat, monkey Human, mouse, rat, monkey, hamster
Target Molecular Weight 45 kDa 45 kDa
Isotype Rabbit IgG Mouse IgG(2b)
Specificity/Sensitivity May cross-react with γ-actin (cytoplasmic isoform)

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