Invitrogen

WWTR1 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

Questions & Answers

Tech Support

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WWTR1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of WWTR1 was performed using 70% confluent log phase HeLa and HT-29 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with WWTR1 Polyclonal Antibody (Product # PA5-52431) at 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of HeLa cells, which is a positive model for WWTR1 expression showing a nuclear and cytoplasmic localization. Panel e represents the merged image of HT-29 cells, that are null for WWTR1 protein expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

WWTR1 Antibody in Immunohistochemistry (IHC) — Immunohistochemical staining of WWTR1 in human kidney using a WWTR1 Polyclonal Antibody (Product # PA5-52431) shows moderate nuclear positivity in glomerular cells and a subset of cells in distal tubules.

WWTR1 Antibody in Western Blot (WB) — Western blot was performed using Anti-WWTR1 Polyclonal Antibody (Product # PA5-52431) and a 55kDa band corresponding to WWTR1 was observed across cell lines tested except HT-29, K-562 and MOLT-4. This product does not show any reactivity to Mouse and Rat tissues. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), Hep G2 (Lane 3), HCT 116 (Lane 4), HT-29 (Lane 5), K-562 (Lane 6), MOLT-4 (Lane 7), Mouse Heart (Lane 8), Rat Heart (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.4 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

WWTR1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. EFO-21 cells were transfected with, control siRNA, WWTR1 siRNA probe #1, and WWTR1 siRNA probe #2 and a decrease in signal intensity was observed in Western Blot application using Anti-WWTR1 Polyclonal Antibody (Product # PA5-52431) used at a dilution of 0.4 µg/mL. Anti-PPIB was used as a loading control. {KD}

Product Details

PA5-52431

Applications	Tested Dilution	Publications
Western Blot (WB)	0.04-0.4 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	1:200-1:500	-
Immunocytochemistry (ICC/IF)	0.25-2 µg/mL	-

Product Specifications
Species Reactivity	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein corresponding to Human WWTR1 View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7.2, with 40% glycerol
Contains	0.02% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2649675

Product Specific Information

Immunogen sequence: MNPKPSSWRK KILPESFFKE PDSGSHSRQS STDSSGGHPG PRLAGGAQHV RSHSSPASLQ LGTGAGAAGS PAQQHAHLRQ QSYDVTDELP LPPGWEMTFT ATGQRYFLNH IEKITTWQDP RKAMNQPLNH MNLHPAVSST

Highest antigen sequence identity to the following orthologs: Mouse - 96%, Rat - 96%.

Target Information

WW domain-containing transcription regulator protein 1 (WWTR1) is a transcriptional coactivator which acts as a downstream regulatory target in the Hippo signaling pathway that plays a pivotal role in organ size control and tumor supression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein STK3/MST2 and STK4/MST1, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1. In turn, MOB1 phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. WWTR1 enhances PAX8 and NKX2-1/TTF1-dependent gene activation. WWTR1 regulates the nuclear accumulation of SMADS and has a key role in coupling them to the transcriptional machinery such as the mediator complex. WWTR1 also regulates embryonic stem-cell self-renewal and promotes cell proliferation and epithelial-mesenchymal transition.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: DKFZp586I1419; FLJ27004; FLJ45718; transcriptional co-activator with PDZ-binding motif; transcriptional coactivator with PDZ binding motif; Transcriptional coactivator with PDZ-binding motif; WW domain-containing transcription regulator protein 1

Gene Aliases: 2310058J06Rik; 2610021I22Rik; C78399; TAZ; WWTR1

UniProt ID: (Human) Q9GZV5, (Mouse) Q9EPK5

Entrez Gene ID: (Human) 25937, (Mouse) 97064

Molecular Function: transcription cofactor