This is a knockout-validated antibody summary, based on the publication "β-Actin specifically controls cell growth, migration, and the G-actin pool" [1] (discussed below), "Relative importance of βcyto- and γcyto-actin in primary mouse embryonic fibroblasts" [2] (see figure 1b for immunocytochemistry and figure 1i for western blot), and "Electron transport chain activity is a predictor and target for venetoclax sensitivity in multiple myeloma" for western blot knockout validation (figure 5a) [3]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Representative immunoblots of cell lysates from ActbL/L and ActbL/L Cre MEFs probed with pan actin (loading control) or beta-actin antibodies. From [1].

Mouse monoclonal IgG1

Company: Sigma

Antibody: beta-actin

Catalog number: AC15

Summary: Mouse monoclonal IgG1 against a slightly modified β-cytoplasmic actin N-terminal peptide conjugated to KLH. Reacts with canine, guinea pig, feline, pig, mouse, chicken, sheep, rabbit, rat, human, and bovine. Suitable for western blot, immunofluorescence, immunohistochemistry, and ELISA.

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Figure 2. Legend: Representative images of ActbL/L (A) and ActbL/L Cre (B) MEFs cultured overnight and costained with antibodies to β-actin (green) and γ-actin (red). Scale bar, 50 μm. From [1].

Western blot | Immunofluorescence

ActbL/L and ActbL/L Cre MEFs. For immunofluorescence, cells were fixed with cold methanol at −20°C for 10 min.

IF: 3% BSA for 1 h.

WB: Fluorescently labeled secondary antibodies.

WB: Odyssey Infrared Imaging System (LI-COR Biosciences, Lincoln, NE).

IF: 20× (NA 0.75), 40× oil (NA 1.35), or 100× oil (NA 1.40) objective on a Delta Vision personalDV microscope using softWoRx 3.7.1 software (Applied Precision).

FITC-conjugated antibody AC15; Abcam (same clone) was used in KO validation by IF.

The same clone (AC-15) is sold as Novus Biologicals NB600-501, NB110-67828; OriGene AM20687PU-N; MilliporeSigma A5441, A3854, A1978, F3022; Santa Cruz Biotechnology sc-69879; LifeSpan Biosciences LS-C171345, LS-C171346, LS-C171347; Abnova MAB8232; Abcam ab6276, ab49900; Invitrogen MA1-91399, AM4302; GeneTex GTX26276; Boster MA1115; Aviva Systems Biology OAAD00224.