Revision 1

#56674Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
CTGF (D8Z8U) Rabbit mAb 86641 20 µl 35 kDa Rabbit IgG
CYR61 (D4H5D) XP® Rabbit mAb 14479 20 µl 41 kDa Rabbit IgG
IGFBP3 (D1U9C) Rabbit mAb 25864 20 µl 40 kDa Rabbit IgG
Integrin β2 (D4N5Z) Rabbit mAb 73663 20 µl 95 kDa Rabbit IgG
Survivin (71G4B7) Rabbit mAb 2808 20 µl 16 kDa Rabbit IgG
Axl (C89E7) Rabbit mAb 8661 20 µl 138 kDa Rabbit IgG
Pan-TEAD (D3F7L) Rabbit mAb 13295 20 µl 50, 53, 55, 60 kDa Rabbit IgG
Lamin B2 (D8P3U) Rabbit mAb 12255 20 µl 68 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The YAP/TAZ Transcriptional Targets Antibody Sampler Kit provides an economical means of detecting proteins whose transcription is subject to regulation by the transcriptional co-activators YAP and/or TAZ. The kit provides enough antibody to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

YAP and TAZ (WWTR1) are transcriptional co-activators that play a central role in the Hippo Signaling pathway that regulates cell, tissue and organ growth. Under growth conditions, YAP and TAZ are translocated to the nucleus, where they interact with DNA-binding transcription factors (e.g., Transcriptional Enhanced Activation Domain [TEAD] proteins) to regulate the expression of genes that control fundamental aspects of cell function, such as proliferation and cell survival (1). A number of genes have been experimentally confirmed as targets of transcriptional regulation by YAP and TAZ. These include the extracellular matrix proteins CTGF, CYR61, and integrin β2 (2-4), the inhibitor of apoptosis protein (IAP) survivin (5), the mechano-sensitive nuclear envelope protein Lamin B2 (6), and the oncogenic receptor tyrosine kinase Axl (7).

  1. Holden, J.K. and Cunningham, C.N. (2018) Cancers (Basel) 10, .
  2. Zhang, H. et al. (2009) J Biol Chem 284, 13355-62.
  3. Chan, S.W. et al. (2011) J Biol Chem 286, 7018-26.
  4. Chan, S.W. et al. (2011) Oncogene 30, 600-10.
  5. Zhang, W. et al. (2015) Cancer Res 75, 4450-7.
  6. Zanconato, F. et al. (2015) Nat Cell Biol 17, 1218-27.
  7. Cui, Z.L. et al. Int J Immunopathol Pharmacol 25, 989-1001.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #56674

    YAP/TAZ Transcriptional Targets Antibody Sampler Kit

    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from HepG2 cells using Pan-TEAD (D3F7L) Rabbit mAb #13295. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from Jurkat cells using Survivin (71G4B7) Rabbit mAb #2808. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 3 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using Axl (C89E7) Rabbit mAb #8661. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using Lamin B2 (D8P3U) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from COS-7 cells transfected with a construct expressing either Myc/DDK-tagged full-length human TEAD1 (hTEAD1-Myc/DDK;+), Myc/DDK-tagged full-length human TEAD2 (hTEAD2-Myc/DDK;+), Myc/DDK-tagged full-length human TEAD3 (hTEAD3-Myc/DDK;+), or Myc/DDK-tagged full-length human TEAD4 (hTEAD4-Myc/DDK;+), using TEAD1 (D9X2L) Rabbit mAb #12292 (A), TEAD2 Antibody #8870 (B), TEAD3 Antibody (C), or Pan-TEAD (D3F7L) Rabbit mAb #13295 (D).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using CYR61 (D4H5D) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, MCF7 cells are negative for CYR61 expression.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from concentrated culture medium of various cell lines using IGFBP3 (D1U9C) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of lysates from Raji (human), BaF3 (mouse) and C6 (rat) cell lines using Survivin (71G4B7E) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 9 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using Integrin β2 (D4N5Z) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of HeLa Cell Extracts, untreated (-) or Axl knock-out (+) using Axl (C89E7) Rabbit mAb, #8661 (upper) or GAPDH (D16H11) XP® Rabbit mAb, #5174 (lower).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from various cell lines using CTGF (D8Z8U) Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower). Asterisk indicates a 120 kDa band of unknown identity detected by this antibody in Western blot analysis.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of lamin B2 from Jurkat cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Lamin B2 (D8P3U) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Lamin B2 (D8P3U) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from COS-7 cells transfected with a construct expressing either Myc/DDK-tagged full-length human TEAD1 (hTEAD1-Myc/DDK;+), Myc/DDK-tagged full-length human TEAD2 (hTEAD2-Myc/DDK;+), Myc/DDK-tagged full-length human TEAD3 (hTEAD3-Myc/DDK;+), or Myc/DDK-tagged full-length human TEAD4 (hTEAD4-Myc/DDK;+), using Pan-TEAD (D3F7L) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 15 Expand Image
    Confocal immunofluorescent analysis of Saos-2 cells, untreated (left) or treated with Brefeldin A #9972 (10 μg/ml, overnight; center), and untreated MCF7 cells (right), using CYR61 (D4H5D) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4804 (fluorescent DNA dye).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 16 Expand Image
    Immunoprecipitation of IGFBP3 protein from HCC1937 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IGFBP3 (D1U9C) Rabbit mAb. Western blot analysis was performed using IGFBP3 (D1U9C) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Survivin siRNA II (+), using Survivin (71G4B7E) Rabbit mAb #2808 and α-Tubulin (11H10) Rabbit mAb #2125. Survivin (71G4B7E) Rabbit mAb confirms silencing of survivin expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of survivin siRNA.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of PANC-1 cells (high-expressing) and MCF7 cells (low-expressing) using CTGF (D8Z8U) Rabbit mAb (green) and an Anti-Na+/K+ ATPase α Mouse mAb (blue). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 20 Expand Image
    Confocal immunofluorescent analysis of HCT 116 cells using Lamin B2 (D8P3U) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 21 Expand Image
    Western blot analysis of extracts from various cell lines using Pan-TEAD (D3F7L) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 22 Expand Image
    Flow cytometric analysis of 293T cells (blue) and PANC-1 cells (green) using CYR61 (D4H5D) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma of the bladder using Survivin (71G4B7E) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 25 Expand Image
    Immunoprecipitation of TEAD proteins from COS-7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Pan-TEAD (D3F7L) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Pan-TEAD (D3F7L) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma showing nuclear localization using Survivin (71G4B7E) Rabbit mAb #2808.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using Axl (C89E7) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Survivin (71G4B7E) Rabbit mAb in the presence of control peptide (left) or Survivin Blocking Peptide #1037 (right).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using Axl (C89E7) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human pituitary adenoma using Survivin (71G4B7E) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Axl (C89E7) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 33 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Survivin (71G4B7E) Rabbit mAb (green). Mitochondria have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded cell pellets, NCI-H1299 (left) or Jurkat (right), using Axl (C89E7) Rabbit mAb.
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 35 Expand Image
    Flow cytometric analysis of untreated Jurkat cells using Survivin (71G4B7E) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 36 Expand Image
    Confocal immunofluorescent analysis of DU 145 (left) and HCC827 (right) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    YAP/TAZ Transcriptional Targets Antibody Sampler Kit: Image 37 Expand Image
    Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and DU145 cells (green, positive) using Axl (C89E7H4) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.