ただ今実施中のプロモーション | 詳細はこちら >>
56612
Hippo Pathway: Upstream Signaling Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Hippo Pathway: Upstream Signaling Antibody Sampler Kit #56612

Reviews ()
Citations (1)
Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 1

Western blot analysis of extracts from control HeLa cells (Lane 1) or YAP knockout HeLa cells (Lane 2) using YAP (D8H1X) XP® Rabbit mAb (upper) or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the YAP knockout HeLa cells confirms the specificity of the antibody for YAP.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and YAP (D8H1X) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across CTGF, a known target gene of YAP (see additional figure containing ChIP-qPCR data).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 3

CUT&RUN was performed with NCI-H2052 cells and YAP (D8H1X) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figures show binding across TSPAN2, a known target gene of YAP (see additional figure containing CUT&RUN-qPCR data).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 4

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and TAZ (D3I6D) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across SMYD3, a known target gene of TAZ (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 5

Immunoprecipitation of Ajuba protein from PANC-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb XP® Isotype Control #3900, and lane 3 is Ajuba (D4D8P) Rabbit mAb. Western blot analysis was performed using Ajuba (D4D8P) Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 6

Western blot analysis of extracts from various cell lines using PTPN14 (D5T6Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 7

Confocal immunofluorescent analysis of PC-3 cells using Merlin (D3S3W) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 8

Western blot analysis of extracts from various cell lines using MST1 (D8B9Q) Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 9

Western blot analysis of extracts from various cell lines using FRMD6 (D8X3R) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 10

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.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 11

Western blot analysis of extracts from various cell lines using YAP (D8H1X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, RL-7 cells are negative for YAP protein expression.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 12

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and YAP (D8H1X) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 6 (upper), including CTGF (lower), a known target gene of YAP (see additional figure containing ChIP-qPCR data).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 13

CUT&RUN was performed with NCI-H2052 cells and YAP (D8H1X) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figures show binding across across chromosome 1 (upper), including TSPAN2 (lower), a known target gene of YAP (see additional figure containing CUT&RUN-qPCR data).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 14

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and either TAZ (D3I6D) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CTGF Promoter Primers #14927, human SMYD3 intron 2 primers, and SimpleChIP® Human CTGF Upstream Primers #14928. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 15

Western blot analysis of extracts from various cell lines using Ajuba (D4D8P) Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 16

Immunoprecipitation of merlin from mouse embryonic fibroblasts (MEF) cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Merlin (D3S3W) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Merlin (D3S3W) Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 17

Western blot analysis of extracts from HeLa cells, serum-starved overnight and then vehicle treated (-) or treated with Staurosporine #9953 (1 μM, 4 hr; +), using MST1 (D8B9Q) Rabbit mAb. Staurosporine induces caspase-mediated cleavage of full-length MST1, yielding a 37 kDa amino-terminal fragment detected by this antibody, and an undetected 18 kDa carboxy-terminal fragment.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 18

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-2052 cells and either YAP (D8H1X) XP® Rabbit mAb, or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CTGF Promoter Primers #14927, human SMYD3 intron 2 primers, and SimpleChIP® Human CTGF Upstream Primers #14928. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 19

CUT&RUN was performed with NCI-H2052 cells and either YAP (D8H1X) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human C1D downstream primers, human TSPAN2 upstream primers, and human ITM2A upstream primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 20

Western blot analysis of extracts from various cells using TAZ (D3I6D) Rabbit mAb (upper) and GAPDH (D16H11) XP® #5174 (lower). As expected, Raji cells are negative for TAZ.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 21

Western blot analysis of extracts from PC-3 cells, and both wild type and Nf2 (-/-) mouse embryonic fibroblasts (MEFs) using Merlin (D3S3W) Rabbit mAb (upper), Ezrin/Radixin/Moesin Antibody #3142 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). (MEF wt and MEF Nf2 (-/-) cells were kindly provided by Dr. Andrea McClatchey and Dr. Andrew Gladden, MGH Cancer Center and Harvard Medical School, Charlestown MA).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 22

Immunoprecipitation of MST1 from Ramos cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or MST1 (D8B9Q) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using MST1 (D8B9Q) Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 23

Flow cytometric analysis of RL-7 cells (blue) and A-204 cells (green) using YAP (D8H1X) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. 

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 24

Confocal immunofluorescent analysis of low confluence MCF 10A cells (left), high confluence MCF 10A (center), and YAP negative RL-7 cells (right) using YAP (D8H1X) XP® Rabbit mAb (green). Blue pseudocolor in lower images = DRAQ5® #4084 (fluorescent DNA dye). Increased nuclear localization of YAP protein is seen in low confluence (proliferating) cells.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin’s lymphoma using YAP (D8H1X) XP® Rabbit mAb performed on the Leica® BOND™ Rx. 

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 26

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using YAP (D8H1X) XP® Rabbit mAb performed on the Leica® BOND™ Rx.  

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 27

Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma using YAP (D8H1X) XP® Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 28

Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma using YAP (D8H1X) XP® Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 29

Immunohistochemical analysis of paraffin-embedded 293T cell pellet, wild-type (left, positive) or YAP/TAZ knockout (right, negative), using YAP (D8H1X) XP® Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded BEN cell pellet, untransfected (left, YAP/TAZ low), YAP-transfected (middle) or TAZ-transfected (right), using YAP (D8H1X) XP® Rabbit mAb (top) or TAZ (E9J5A) XP® Rabbit mAb #72804 (bottom).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 31

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma (left), colon carcinoma case 1 (middle) or colon carcinoma case 2 (right), using YAP (D8H1X) XP® Rabbit mAb (top) or TAZ (E9J5A) XP® Rabbit mAb #72804 (bottom).

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 32

Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using YAP (D8H1X) XP® Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 33

Immunohistochemical analysis of paraffin-embedded PANC-1 (left) and RL-7 (right) cell pellets using YAP (D8H1X) XP® Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 34

Immunohistochemical analysis of paraffin-embedded human benign prostatic hyperplasia using YAP (D8H1X) XP® Rabbit mAb.

Hippo Pathway: Upstream Signaling Antibody Sampler Kit: Image 35

Immunoprecipitation of YAP protein from A-204 cell extracts using Rabbit (DA1E) mAb XP® Isotype Control #3900 (lane 2) or YAP (D8H1X) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using YAP (D8H1X) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.

To Purchase # 56612T
Product # Size Price Inventory
56612T
1 Kit  (7 x 20 µl)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
YAP (D8H1X) XP® Rabbit mAb 14074 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
  • C&R
Hm H Mk M R 65-78 Rabbit IgG
TAZ (D3I6D) Rabbit mAb 70148 20 µl
  • WB
  • IP
  • ChIP
H M 50 Rabbit IgG
Ajuba (D4D8P) Rabbit mAb 34648 20 µl
  • WB
  • IP
H Mk 55 Rabbit IgG
PTPN14 (D5T6Y) Rabbit mAb 13808 20 µl
  • WB
H M R 160 Rabbit IgG
Merlin (D3S3W) Rabbit mAb 12888 20 µl
  • WB
  • IP
  • IF
Hm H Mk M R 70 Rabbit IgG
MST1 (D8B9Q) Rabbit mAb 14946 20 µl
  • WB
  • IP
H M R 60 Rabbit IgG
FRMD6 (D8X3R) Rabbit mAb 14688 20 µl
  • WB
  • IP
H M R 78 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Hippo Pathway Proteins Antibody Sampler Kit provides an economical means of detecting proteins that have been identified as upstream regulators of the Hippo Signaling Pathway. The kit provides enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Hippo Pathway Proteins Antibody Sampler Kit detects endogenous levels of its target protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ala200 of mouse TAZ, Phe608 of human FRMD6, Leu737 of human PTPN14, Gln470 of human merlin, Arg77 of human Ajuba, Pro9 of human MST1, and recombinant protein corresponding to the carboxy terminus of human YAP protein. The sequence region surrounding the YAP epitope is 100% conserved among all known isoforms of YAP.

Background

The Hippo Signaling Pathway is an evolutionarily conserved signaling cascade that regulates cell, tissue and organ growth in response to diverse environmental cues, including cell density, mechanical stress, growth factors and cytokines, and metabolic signaling (1,2). In mammals, the core components of this pathway are YAP and TAZ, transcriptional co-activators that regulate the expression of target genes that control cell proliferation and survival (3). The transcriptional control of YAP/TAZ target genes is regulated through a complex kinase cascade involving mammalian sterile-20-like kinases (MST1/2), LATS1/2 kinases and key adaptor proteins. Phosphorylation of YAP and TAZ by LATS kinases results in cytoplasmic sequestration of YAP and TAZ, inhibiting transcriptional activation of their target genes. A number of proteins have been identified that function upstream of MST1/2 and LATS1/2 to influence LATS-mediated phosphorylation of YAP/TAZ. These include the tumor suppressor protein Merlin (4), the FERM-domain containing protein 6 (FRMD6/Willin) (5), the LIM-domain containing adaptor protein Ajuba (6,7), and the protein tyrosine phosphatase PTPN14 (8).

  1. Meng, Z. et al. (2016) Genes Dev 30, 1-17.
  2. Watt, K.I. et al. (2017) Front Physiol 8, 942.
  3. Fu, V. et al. (2017) Curr Opin Cell Biol 49, 99-107.
  4. Su, T. et al. (2017) Dev Cell 40, 478-490.
  5. Angus, L. et al. (2012) Oncogene 31, 238-50.
  6. Das Thakur, M. et al. (2010) Curr Biol 20, 657-62.
  7. Reddy, B.V. and Irvine, K.D. (2013) Dev Cell 24, 459-71.
  8. Liu, X. et al. (2013) Oncogene 32, 1266-73.

Pathways & Proteins

Explore pathways + proteins related to this product.

使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.