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3019
Met Signaling Antibody Sampler Kit

Met Signaling Antibody Sampler Kit #3019

Western Blotting Image 1

Western blot analysis of extracts from HEK293 cells, untreated or treated with In1B for indicated times, using Phospho-Gab1 (Tyr307) Antibody (upper) or Gab1 Antibody #3232 (lower).

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Western Blotting Image 2

Western blot analysis of extracts from HEK293, 3T3-L1, C6 and COS cells, using Gab1 Antibody.

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Western Blotting Image 3

Western blot analysis of extracts from A431 cells, untreated or HGF-treated using Phospho-Met (Tyr1003) (13D11) Rabbit mAb (upper and middle) and Met (25H2) Mouse mAb #3127 (lower). The middle blot was treated with CIP phosphatase before antibody probing.

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Western Blotting Image 4

Western blot analysis of cell extracts from HeLa cells, untreated or stimulated with HGF, using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (upper) and Met (25H2) Mouse mAb #3127 (lower).

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Western Blotting Image 5

Western blot analysis of cell lysates of H4IIE cells untreated or treated with HGF, using Phospho-Met (Tyr1349) (130H2) Rabbit mAb (upper), or Met (25H2) Mouse mAb #3127 (lower).

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Western Blotting Image 6

Western blot analysis of extracts from HT-29 (Met+), SK-BR-3 (Met-), and T-47D (Met-) cells using Met (D1C2) XP® Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).

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Western Blotting Image 7

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.

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IP Image 8

Immunoprecipitation of Gab 1 from insulin treated Cos cell extracts using Gab 1 antibody (Lane 1). Lane 2: No antibody control. Lane 3: Input control.

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Western Blotting Image 9

Western blot analysis of purified active Ron kinase using a Phospho-Ron (Ser1394) Antibody (A), a Phospho-Ron (Tyr1238/1239) Antibody (B), Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (C) and Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (D). This demonstrates that Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb does not cross-react with phospho-Ron by western analysis.

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human metastatic lung carcinoma using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 11

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded xenografts from 3T3-Met (left) and 3T3-Ron cells (right) using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb, indicating that this antibody does not cross-react with activated Ron by immunohistochemistry. Image courtesy of Pfizer, Inc.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human papillary renal cell carcinoma using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis on Src-transfected NIH/3T3 cells, using a Phospho-Src Family (Tyr416) Antibody (left) or Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (right), indicating that the antibody does not cross-react with Src phosphorylated at Tyr416 via immunohistochemistry.

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IHC-P (paraffin) Image 16

Immunohistochemical analysis of paraffin-embedded cell pellets, MKN-45 (left) and T-47D (right), using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 17

Immunohisochemical analysis of paraffin-embedded HCC827 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.

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IHC-F (frozen) Image 18

Immunohistochemical analysis of frozen MKN-45 xenograft using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 19

Immunohistochemical analysis using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb on SignalSlide™ Phospho-Met (1234/1235) IHC Controls #8118 [MKN45 cells, untreated (left) or SU11274-treated (right)].

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Flow Cytometry Image 20

Flow cytometric analysis of T-47D cells (blue) and HT-29 cells (green) using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded papillary renal cell carcinoma using Phospho-Met (Tyr1234/1225) (D26) XP® Rabbit mAb.

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IF-IC Image 22

Confocal immunofluorescent analysis of HT-29 and T-47D cells using Met (D1C2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-F (frozen) Image 23

Immunohistochemical analysis of frozen MKN45 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.

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Flow Cytometry Image 24

Flow cytometric analysis of MKN-45 cells, untreated (green) or treated with SU11274 (blue).

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IF-IC Image 25

Confocal immunofluorescence analysis of MKN45 cells, untreated (left) or treated with SU11274 (1 μM, 3 hours; right), using Phospho-MET (Tyr1234/Tyr1235) (D26) XP® Rabbit mAb. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Gab1 (Tyr307) Antibody 3234 20 µl
  • WB
H M 115 Rabbit 
Gab1 Antibody 3232 20 µl
  • WB
  • IP
H M R Mk 110 Rabbit 
Phospho-Met (Tyr1003) (13D11) Rabbit mAb 3135 20 µl
  • WB
H M R 145 Rabbit IgG
Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb 3077 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 145 Rabbit 
Phospho-Met (Tyr1349) (130H2) Rabbit mAb 3133 20 µl
  • WB
H M R 145 Rabbit 
Met (D1C2) XP® Rabbit mAb 8198 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H 140, 170 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Met Signaling Antibody Sampler Kit provides an economical means to investigate Met signaling. The kit contains primary and secondary antibodies to perform two western blots with each antibody.

Each antibody in the Met Signaling Antibody Sampler Kit recognizes endogenous levels of its specific target and does not cross-react with other family members unless otherwise indicated. Phospho-Met (Tyr1234/1235) (D26) Rabbit mAb may cross-react with overexpressed tyrosine phosphorylated Src proteins in Western blot. Phospho-Met (Tyr1349) (130H2) Rabbit mAb may cross-react with other activated protein tyrosine kinases. Phospho-Met (Tyr1003) (13D11) Rabbit mAb may cross-react with other activated protein tyrosine kinases. Phospho-Gab1 (Tyr307) Antibody cross-reacts with phosphorylated Gab2 and potentially with phosphorylated Gab3.

Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides or peptides corresponding to residues surrounding: Tyr1003, Tyr1234/1235, Tyr1349, or near the carboxy terminus of human Met protein. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide or peptide corresponding to residues surrounding Tyr472 and Tyr307 of human Gab1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Met, a high affinity tyrosine kinase receptor for hepatocyte growth factor (HGF, also known as scatter factor), is a disulfide-linked heterodimer made of 45 kDa α- and 145 kDa β-subunits (1,2). The α-subunit and the amino-terminal region of the β-subunit form the extracellular domain. The remainder of the β-chain spans the plasma membrane and contains a cytoplasmic region with tyrosine kinase activity. Interaction of Met with HGF results in autophosphorylation at multiple tyrosines, which recruit several downstream signaling components, including Gab1, c-Cbl and PI3 kinase (3). These fundamental events are important for all of the biological functions involving Met kinase activity. Addition of a phosphate at cytoplasmic Tyr1003 is essential for ubiquitination and Met protein degradation (4). Phosphorylation of Tyr1234/1235 in the Met kinase domain is critical to kinase activation. Phosphorylation of Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon and breast cancers. Thus, Met is an attractive cancer therapeutic and diagnostic target (6).

  1. Weidner, K.M. et al. (1993) Mol Immunol 30, 1003-11.
  2. Park, M. et al. (1986) Cell 45, 895-904.
  3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
  4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
  5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
  6. Traxler, P. et al. (2001) Med Res Rev 21, 499-512.
Entrez-Gene Id
2549 , 4233
Swiss-Prot Acc.
Q13480 , P08581
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.

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