This is a knockout-validated antibody summary, based on the publication "Whole animal knockout of smooth muscle alpha-actin does not alter excisional wound healing or the fibroblast-to-myofibroblast transition", as cited below [1], "Abnormal angiogenesis of placenta in progranulin‑deficient mice" for western blot knockout validation (figure 4e) [2] and "Repression of Osmr and Fgfr1 by miR-1/133a prevents cardiomyocyte dedifferentiation and cell cycle entry in the adult heart" for western blot knockout validation (figure s10f) [3]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG2a

Company: Sigma-Aldrich

Antibody: Actin, α-Smooth Muscle

Catalog number: A2547

Summary: Mouse monoclonal IgG2a against an N-terminal synthetic decapeptide of α-smooth muscle actin. Reacts with rabbit, guinea pig, mouse, chicken, snake, sheep, goat, human, frog, rat, canine, and bovine. Suitable for western blot, immunofluorescence and immunohistochemistry (paraffin and frozen).

Western blot | Immunocytochemistry

WT and SMαA-null (SMαA−/−) primary dermal fibroblasts from mice cultured for 4 days in complete medium with 1 ng/ml TGF-β1 (Sigma-Aldrich) to promote myofibroblast differentiation. WB: Whole cell lysates were collected in 1% SDS lysis buffer.

WB: Blocking Buffer (Rockland Immunochemicals, Inc., Gilbertsville, PA).

WB: 1:5,000 dilution in Blocking Buffer overnight at 4°C.

IC: 1:1,000 dilution.

IC: 1:3,000 dilution goat anti-mouse IgG conjugated to horseradish peroxidase (BioRad, Hercules, CA) at ambient temperature for 1 hour.

WB: ECL Western Blotting Substrate (Pierce, Rockford, IL) and a BioSpectrum 600 Imaging System (UVP, Upland, CA).

IC: Fluorescence microscopy and image capture was performed using an Olympus Provis AX-70 photomicroscope (Olympus) equipped with a QImaging digital camera (QImaging, Surrey, British Columbia, Canada).

Immunofluorescence and immunoblot of WT and SMαA-null (SMαA−/−) fibroblasts with anti-SMαA antibody. Fibroblasts were cultured with TGF-β1 for 4 days to promote myofibroblast formation. DAPI was used to detect nuclei. Bar=100 μm. Please see Figure 2a and 5a in the article [1].

The same clone (1A4) is sold as Bio-Rad MCA5781GA; BioLegend 904601; OriGene TA327604; MilliporeSigma C6198, A2547, F3777, A5228, A5691; Santa Cruz Biotechnology sc-32251, sc-32251 AC; LifeSpan Biosciences LS-C86744, LS-C87562, LS-C87563, LS-C95300, LS-C124868, LS-B5147, LS-C171306, LS-C182754, LS-C182755, LS-C187560, LS-C202806; Abnova MAB1492; Abcam ab7817, ab8211, ab184675, ab125057, ab202368, ab203696; R&D Systems MAB1420-SP, IC1420T-025, IC1420V-025, MAB1420, IC1420P, IC1420A, IC1420G, IC1420S-025, IC1420C, IC1420T-100, IC1420N, IC1420V-100, IC1420R-100UG; Invitrogen MA1-06110, 14-9760-80, 14-9760-82, 41-9760-80, 41-9760-82, 50-9760-80, 50-9760-82, 53-9760-80, 53-9760-82, 14-9760-95, 14-9760-37; MyBioSource MBS175016; Boster MA1106; Fitzgerald Industries 10R-6761.