This is a knockout-validated antibody summary, based on the three publications "The steroid receptor coactivator-3 is required for developing neuroendocrine tumor in the mouse prostate" [1], "Tead and AP1 Coordinate Transcription and Motility" [2], and "Knockout of SRC-1 and SRC-3 in Mice Decreases Cardiomyocyte Proliferation and Causes a Noncompaction Cardiomyopathy Phenotype" [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. Immunohistochemistry for SRC-3 (brown) in the prostate sections prepared from 8-week-old floxed (3FloxT) and specifically ablated in AR+/Syp- prostatic epithelial cells (3PEKOT) mice. From [1].

Antibody: SRC-3

Catalog number: 2126

Summary: Rabbit monoclonal raised against a synthetic peptide corresponding to the sequence of human SRC-3 protein. Detects endogenous levels of total SRC-3 protein (all isoforms) from human, mouse, rat, and monkey. Used for western blotting, immunoprecipitation, and chromatin IP.

Company: Cell Signaling

IHC, immunohistochemistry

Fixed prostate tissues sections (paraffin embeded) prepared from 8-week-old floxed (3FloxT) and specifically ablated in AR+/Syp- prostatic epithelial cells (3PEKOT) mice.

Biotinylated anti-Rabbit IgG (Vector labs) used at 1:400 dilution

Signal was enhanced using the VECTASTAIN ABC system and visualized with DAB kit.

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Figure 2. Immunoblot analysis of SRC3 in HCT116 cells with or without expression of Crispr knockout constructs against SRC1, SRC2 and SRC3 (SRC1-3 KO). GAPDH was used as loading control. From [2].

Western blot

Control and SRC1-3 KO HCT116 cells.

HRP conjugated secondary antibodies (Jackson Laboratories).

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Figure 3. Western blot analysis of SRC-1 and SRC-3 protein levels in WT, S1KO and S3KO hearts at P0 (positive, SRC-1 knockout and SRC-3 knockout controls, respectively) and SRC-3f/f, SRC-3d/d, S1KO;SRC-3f/f and S1KO;SRC-3d/d primary cardiomyocytes. Each pool of the primary cardiomyocytes was prepared from multiple hearts with the same genotype. α-tubulin served as a loading control. The lower band detected by SRC-3 antibody is a SRC-3 splicing isoform. From [3].

Western blot

WT, SRC-1 knockout and SRC-3 knockout hearts at P0, SRC-3f/f, SRC-3d/d, S1KO;SRC-3f/f and S1KO;SRC-3d/d primary cardiomyocytes. Frozen tissue powders and harvested cells were extracted with RIPA buffer containing 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1% NP-40 and 0.25% Na-deoxycholate.

Appropriate secondary antibodies conjugated with horseradish peroxidase.

ECL reagent kit.

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.