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8579
Hippo Signaling Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Hippo Signaling Antibody Sampler Kit #8579

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Hippo Signaling Antibody Sampler Kit: Image 1

Western blot analysis of extracts from Hep G2 cells, untreated (-) or λ-phosphatase-treated (+), using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). YAP protein isoform 1 Ser397 corresponds to Ser381 of YAP isoform 2, as reported by Zhao et al. (2010) Genes Dev 24, 72-85 (9).

Hippo Signaling Antibody Sampler Kit: Image 2

Western blot analysis of extracts from various cell lines using LATS1 (C66B5) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 3

Western blot analysis of extracts from MCF7 cells, either untreated (-) or treated (+) with H2O2 (2.5 mM, 30 min) and Ramos cells, either untreated (-) or treated (+) with λ phosphatase, using Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb (upper) and MOB1 Antibody #3863 (lower).

Hippo Signaling Antibody Sampler Kit: Image 4

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 5

Western blot analysis of extracts from various cell lines using MOB1 (E1N9D) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 6

Immunoprecipitation of MOB1 protein from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or MOB1 (E1N9D) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using MOB1 (E1N9D) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 7

Western blot analysis of extracts from various cell lines using MST1 Antibody.

Hippo Signaling Antibody Sampler Kit: Image 8

Western blot analysis of extracts from various cell lines using MST2 Antibody.

Hippo Signaling Antibody Sampler Kit: Image 9

Confocal immunofluorescent analysis of HCT 116 (higher expressing, left) and ACHN (lower expressing, right) cells using SAV1 (D6M6X) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Hippo Signaling Antibody Sampler Kit: Image 10

Western blot analysis of extracts from HeLa cells (lane 1) or YAP knock-out cells (lane 2) using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb #13008 (upper), and GAPDH (D6H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the YAP knock-out HeLa cells confirms specificity of the antibody for YAP.

Hippo Signaling Antibody Sampler Kit: Image 11

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma, control (left) or λ-phosphatase treated (right), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 12

Western blot analysis of extracts from various cell lines using YAP/TAZ (D24E4) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 13

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 Signaling Antibody Sampler Kit: Image 14

Western blot analysis of extracts from MDA-MB-231 cells, vehicle treated (-) or treated with epinephrine (10 μM, 60 min; +) or Forskolin #3828 (10 μm, 60 min; +), using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8547 (lower). YAP protein isoform 1 Ser397 corresponds to Ser381 of YAP isoform 2, as reported by Zhao, B. et al. (2010) Genes Dev 24, 72-85 (9).

Hippo Signaling Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma control (left) or lambda phosphatase-treated (right) using Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 16

Western blot analysis of extracts from HeLa cells treated with Staurosporine #9953 for the indicated times using MST2 Antibody.

Hippo Signaling Antibody Sampler Kit: Image 17

Immunoprecipitation of SAV1 protein from A-204 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or SAV1 (D6M6X) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using SAV1 (D6M6X) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 18

Immunohistochemical analysis of paraffin-embedded cell pellets, A-204 (left) and RL-7 (right), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 19

Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 20

Immunohistochemical analysis of paraffin-embedded MCF7 cell pellets, control (left) or H2O2-treated (right), using Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb.

Hippo Signaling Antibody Sampler Kit: Image 21

Western blot analysis of extracts from various cell lines using SAV1 (D6M6X) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Hippo Signaling Antibody Sampler Kit: Image 22

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Hippo Signaling Antibody Sampler Kit: Image 23

Schematic diagram showing annotation of the amino acid sequence surrounding Ser397 in different human YAP isoforms. Asterisk (*) indicates annotation of the phosphorylation site as described in Zhao et al. 2010 [9].

Hippo Signaling Antibody Sampler Kit: Image 24

Western blot analysis of extracts from HeLa cells (lane 1) or YAP knock-out cells (lane 2) using YAP/TAZ (D24E4) Rabbit mAb #8418 (upper), and GAPDH (D6H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the YAP knock-out HeLa cells confirms specificity of the antibody for YAP.

Hippo Signaling Antibody Sampler Kit: Image 25

Western blot analysis of extracts from PANC-1 cells, untreated (-) or λ-phosphatase-treated (+), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Hippo Signaling Antibody Sampler Kit: Image 26

Western blot analysis of MDA-MB-231 cells, vehicle-treated (-) or treated with Forskolin #3828 (10 μM, 60 min; +) or epinephrine (10 μM, 60 min; +), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). Note the induction of YAP (Ser127) phosphorylation after treatment with forskolin or epinephrine, consistent with the findings reported in Xu et al. (2012) [9].

Hippo Signaling Antibody Sampler Kit: Image 27

Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.

To Purchase # 8579T
Product # Size Price
8579T
1 Kit  (9 x 20 µl) $ 609

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb 13619 20 µl
  • WB
  • IP
H M R 65-78 Rabbit IgG
LATS1 (C66B5) Rabbit mAb 3477 20 µl
  • WB
  • IP
H Mk M 140 Rabbit IgG
Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb 8699 20 µl
  • WB
  • IHC
H Mk M R 24 Rabbit IgG
MOB1 (E1N9D) Rabbit mAb 13730 20 µl
  • WB
  • IP
Hm H Mk M R 24 Rabbit IgG
MST1 Antibody 3682 20 µl
  • WB
  • IP
B H Mk M R 59 Rabbit 
MST2 Antibody 3952 20 µl
  • WB
  • IP
B H Mk M R 60 Rabbit 
SAV1 (D6M6X) Rabbit mAb 13301 20 µl
  • WB
  • IP
  • IF
H M 45 Rabbit IgG
Phospho-YAP (Ser127) (D9W2I) Rabbit mAb 13008 20 µl
  • WB
  • IP
  • IHC
H M R 65-78 Rabbit IgG
YAP/TAZ (D24E4) Rabbit mAb 8418 20 µl
  • WB
  • IP
H Mk M 55, 78 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Hippo Signaling Antibody Sampler Kit provides an economical means of detecting target proteins of the Hippo signaling pathway. The kit contains enough primary antibody to perform two western blots per primary.

Specificity / Sensitivity

Phospho-YAP (Ser397) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser397. This residue corresponds to Ser381 of YAP isoform 2, as reported by Zhao, B. et al. (2010) Genes Dev 24, 72-85 (9). Phospho-YAP (Ser127) (D9W2I) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser127. YAP/TAZ (D24E4) Rabbit mAb recognizes endogenous levels of total YAP and TAZ proteins. LATS1 (C66B5) Rabbit mAb recognizes endogenous levels of total LATS1 protein. Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb recognizes endogenous levels of MOB1 protein only when phosphorylated at Thr35. MOB1 (E1N9D) Rabbit mAb recognizes endogenous levels of total MOB1 protein. This antibody detects both MOB1A and MOB1B. Mst1 Antibody recognizes endogenous levels of total Mst1 protein. This antibody does not cross-react with Mst2-4. Mst2 Antibody recognizes endogenous levels of total Mst2 protein. This antibody does not cross-react with Mst1, Mst3, or Mst4. SAV1 (D6M6X) Rabbit mAb recognizes endogenous levels of total SAV1 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Mst1 protein, or residues near the amino terminus of human Mst2 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human MOB1A protein, residues surrounding Gly180 of human LATS1 protein, residues near the carboxy terminus of human SAV1 protein, or residues surrounding Asp362 of human TAZ protein. Phospho-specific monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr35 of human MOB1 protein, residues surrounding Ser397 of human YAP protein isoform 1, or residues surrounding Ser127 of human YAP protein.

Background

Hippo signaling is an evolutionarily conserved pathway that controls cell proliferation, apoptosis, and organ size in response to changing cell density levels (1,2). At relative low cell density, transcription co-activators YAP and TAZ bind transcription factors to induce expression of genes that favor cell growth and proliferation. As cell density increases, interaction between membrane-bound upstream hippo pathway regulators trigger activation of cytoplasmic kinases Mst1/2 and LATS1/2. Activated Mst kinase (the eponymous Hippo in Drosophila) associates with the adaptor Sav1 and phosphorylates MOB1 to activate LATS kinase, which phosphorylates YAP and TAZ to suppress cell proliferation (3).

  1. McNeill, H. and Woodgett, J.R. (2010) Nat Rev Mol Cell Biol 11, 404-13.
  2. Zeng, Q. and Hong, W. (2008) Cancer Cell 13, 188-92.
  3. Zhao, B. et al. (2007) Genes Dev 21, 2747-61.

Limited Uses

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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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.