This is a knockout-validated antibody summary, based on the publications "Genome-wide CRISPR screening reveals genetic modifiers of mutant EGFR dependence in human NSCLC" (Western blot, figure 2a in the article) [2] and "YAP/TAZ initiate and maintain Schwann cell myelination", as cited below [1], "Axon-dependent expression of YAP/TAZ mediates Schwann cell remyelination but not proliferation after nerve injury" for western blot knockout validation (see [3], and "Alternative splicing reverses the cell-intrinsic and cell-extrinsic pro-oncogenic potentials of YAP1" for western blot knockout validation (figure 2a) [4]. 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 of sciatic nerve lysates from P60 mice of the indicated genotypes, showing efficient, selective deletion of YAP in Yap cKO, TAZ in Taz cKO, and both YAP/TAZ in Yap/Taz cDKO. Blots in the left panels were exposed for longer than the blot in the right panels. (B) Immunohistochemical analysis showing Schwann cells (SC) nuclei (red) of Yap cKO and Taz cKO that retain YAP/TAZ immunostaining (green) because SCs express both YAP and TAZ. YAP/TAZ are completely eliminated from most cDKO SCs (arrowheads); occasional SCs exhibit low or faint YAP/TAZ immunoreactivity (for example, arrow). Asterisks indicate non-Schwann cells with YAP/TAZ immunostaining. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: YAP/TAZ

Catalog number: 8418

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Asp362 of human TAZ protein. Reacts with human, mouse and monkey. Suitable for western blot, immunoprecipitation and immunohistochemistry (paraffin).

Western blot | Immunohistochemistry

Sciatic nerves from WT, Yap-KO, Taz-KO and Yap/Taz-cDKO mice. WB: sciatic nerves were lysed in RIPA buffer (25 mM Tris pH 7.5, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, 1 mM EDTA, 0.1% SDS), and for immunoprecipitation, lysis was in co-IP buffer (50 mM Tris pH 8.0, 150 mM NaCl, 0.5% Triton X-100, 10% glycerol, 1 mM EDTA), both containing protease and phosphatase inhibitors. IHC: sciatic nerves were removed and immediately fixed in 4% paraformaldehyde/phosphate buffer for 1 hr for cryosectioning.

IHC: 2% BSA/ PBS.

WB: 1:1,000 dilution.

IHC: 1:200 dilution.

WB: 1:5,000 dilution HRP-linked donkey anti-rabbit (GE Healthcare, United Kingdom) or 1:20,000 HRP-linked mouse anti-rabbit, light-chain specific (Jackson ImmunoResearch, West Grove, PA, 211-032-171).

WB: Blots were developed using ECL, ECL Plus (GE Healthcare) or Odyssey (LiCor).

IHC: Conventional fluorescence microscopy was performed using Olympus BX53 and Zeiss Axio Imager two microscopes. Images were captured using Hamamatsu ORCA R2 C10600 or Zeiss Axiocam HRm Rev three digital cameras, respectively, and Metamorph software.

Rabbit anti-phospho-YAP (Cell Signaling D9W2I #13008; 1:200) and mouse anti-YAP1 (Abcam ab56701; 1: 200) are also KO validated by IHC (Fig. 1 S1).