This is a knockout-validated antibody summary, based on the publication "A genome editing vector that enables easy selection and identification of knockout cells", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. (A) Western blotting analysis of each knockout cell line using an antibody specific for each actin isoform. The lane indicated as Cont was loaded with whole cell lysate of the parental cell line, U2OS. Beta-tubulin was used as the internal control for Western blotting. (B) Immunofluorescence of knockout cell lines. Cells were stained with the same isoform-specific antibodies as those used in Western blotting. Pseudo color is not consistent with the fluorescent dye color of the secondary antibodies. All images are shown with the same magnification. Bar: 100 um. Please see Figure 4 in the article [1].

Mouse monoclonal IgG2a

Company: Sigma-Aldrich

Antibody: β-Actin

Catalog number: A2228

Summary: Mouse monoclonal IgG2a against a slightly modified β-cytoplasmic actin N-terminal peptide. Reacts with canine, guinea pig, rabbit, bovine, chicken, rat, sheep, carp, Drosophila, pig, cat, mouse, human. Suitable for western blot, immunocytochemistry, immunohistochemistry and microarray.

Western blot | Immunocytochemistry

Parental and β-Actin-KO U2OS cells.

WB: 1:2,000 dilution.

WB: 1:3,000 dilution horseradish peroxidase–conjugated anti-mouse antibody (KPL, Gaithersburg, MD).

IC: anti-mouse IgG2a conjugated with Alexa Fluor 594 (Invitrogen).

WB: the chemiluminescence signal (Western BLoT Hyper HRP Substrate, Takara-Bio) was detected with a C-DiGit Blot Scanner (LiCor, Lincoln, NE).

IC: conventional fluorescence microscope (IX-81, Olympus, Tokyo, Japan) equipped with an Orca Flash2.8 C-MOS camera (Hamamatsu Photonics, Hamamatsu, Japan).