Actin, Smooth Muscle antibody | knockout validation | Sigma-Aldrich A7607

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]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Mouse monoclonal IgG2b

Company: Sigma-Aldrich

Antibody: Actin, Smooth Muscle

Catalog number: A7607

Summary: Mouse monoclonal IgG2b against a partially purified chicken gizzard actin. Reacts with chicken, monkey, human, rat, and mouse. Suitable for western blot, immunocytochemistry, immunohistochemistry and ELISA.

Validation Method

Western blot

Sample

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.

Blocking agent

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

Primary incubation

WB: 1:300 dilution dilution in Blocking Buffer overnight at 4°C.

Detection

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

Figure

Immunoblot of WT and SMαA-null (SMαA−/−) fibroblasts with clone CGA7 antibody to determine SMγA and SMαA levels. Arrow indicates molecular weight of actin. Fibroblasts were cultured with TGF-β1 for 4 days to promote myofibroblast formation. Please see Figure 5b in the article [1].

Clone note

The same clone (CGA7) is sold as Santa Cruz Biotechnology sc-53015; Sigma-Aldrich A7607; EMD Millipore MAB1522.

References
  1. Tomasek J, Haaksma C, Schwartz R, Howard E. Whole animal knockout of smooth muscle alpha-actin does not alter excisional wound healing or the fibroblast-to-myofibroblast transition. Wound Repair Regen. 2013;21:166-76 pubmed publisher