This is a knockout-validated antibody summary, based on the publication "Sestrins function as guanine nucleotide dissociation inhibitors for Rag GTPases to control mTORC1 signaling" [1] (discussed below), "TSC2 mediates hyperosmotic stress-induced inactivation of mTORC1" [2] (discussed below), "Lysosomal recruitment of TSC2 is a universal response to cellular stress" [3] (see supplementary figure 10a in the article), "Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling" [4] (see figure 1a in the article), "RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death" (Figure 1b, Western blot) [5], and "3,4-Dimethoxychalcone induces autophagy through activation of the transcription factors TFE3 and TFEB" for western blot knockout validation (figure 4j) [6], "TSC2 regulates lysosome biogenesis via a non-canonical RAGC and TFEB-dependent mechanism" for western blot knockout validation (figure 3e) [7] and "Poly-L-arginine promotes asthma angiogenesis through induction of FGFBP1 in airway epithelial cells via activation of the mTORC1-STAT3 pathway" for western blot knockout validation (figure 3g) [8]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Total cell lysates from Tsc2+/+ and Tsc2−/− mouse embryonic fibroblasts (MEFs) were analyzed by immunoblotting. Actin was used as loading control. From [1]

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: TSC2

Catalog number: 4308

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to carboxy-terminal residues of human tuberin protein. Reacts with human, mouse, rat, hamster, and monkey. Suitable for western blot, immunoprecipitation, immunofluorescence/immunocytochemistry, and flow cytometry.

Western blot

Tsc2+/+ and Tsc2−/− mouse embryonic fibroblasts. Cells were lysed in 1X Cell Lysis Buffer (CST) supplemented with one tablet of EDTA-free protease inhibitor (Roche) per 50 ml and one tablet of PhosSTOP (Roche) per 10 ml.

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Figure 2. Immunoblot of TSC2 in control and TSC2 knock-out MEFs. Tubulin was used as loading control. From [2].

Western blot

Control and TSC2 knock-out MEFs. Cells were lysed in-well with ice-cold Triton lysis buffer (50 mM Tris pH 7.5, 1% Triton X-100, 150 mM NaCl, 50 mM NaF, 2 mM Na-vanadate, 0.011 gr/ml beta-glycerophosphate, 1× PhosSTOP phosphatase inhibitors and 1× Complete protease inhibitors) for 10 minutes on ice.

1:2,000 dilution.

LICOR Fc detection system.