Revision 1

#8579Store at -20C

1 Kit

(9 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
Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb 13619 20 µl 65-78 kDa Rabbit IgG
LATS1 (C66B5) Rabbit mAb 3477 20 µl 140 kDa Rabbit IgG
Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb 8699 20 µl 24 kDa Rabbit IgG
MOB1 (E1N9D) Rabbit mAb 13730 20 µl 24 kDa Rabbit IgG
MST1 Antibody 3682 20 µl 59 kDa Rabbit 
MST2 Antibody 3952 20 µl 60 kDa Rabbit 
SAV1 (D6M6X) Rabbit mAb 13301 20 µl 45 kDa Rabbit IgG
Phospho-YAP (Ser127) (D9W2I) Rabbit mAb 13008 20 µl 65-78 kDa Rabbit IgG
YAP/TAZ (D24E4) Rabbit mAb 8418 20 µl 55, 78 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 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.

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

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.

Background References

    Trademarks and Patents

    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.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Revision 1
    #8579

    Hippo Signaling Antibody Sampler Kit

    Hippo Signaling Antibody Sampler Kit: Image 1 Expand Image
    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 2 Expand Image
    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 3 Expand Image
    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 4 Expand Image
    Western blot analysis of extracts from various cell lines using MOB1 (E1N9D) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using LATS1 (C66B5) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using MST1 Antibody.
    Hippo Signaling Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using MST2 Antibody.
    Hippo Signaling Antibody Sampler Kit: Image 8 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.
    Hippo Signaling Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using YAP/TAZ (D24E4) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 10 Expand Image
    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 11 Expand Image
    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 12 Expand Image
    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 13 Expand Image
    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 14 Expand Image
    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 15 Expand Image
    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 16 Expand Image
    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 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-MOB1 (Thr35) (D2F10) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 18 Expand Image
    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 19 Expand Image
    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 20 Expand Image
    Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 21 Expand Image
    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 22 Expand Image
    Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 23 Expand Image
    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 Expand Image
    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 25 Expand Image
    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 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded cell pellets, A-204 (left) and RL (right), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.
    Hippo Signaling Antibody Sampler Kit: Image 27 Expand Image
    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).