Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Met Signaling Antibody Sampler Kit #3019

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-Gab1 (Tyr307) Antibody #3234 40 µl W H M 115 Rabbit
Gab1 Antibody #3232 40 µl W IP H M R Mk 110 Rabbit
Phospho-Met (Tyr1003) (13D11) Rabbit mAb #3135 40 µl W H M R 145 Rabbit IgG
Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb #3077 40 µl W IP IHC-P IHC-F IF-IC F H M R 145 Rabbit
Phospho-Met (Tyr1349) (130H2) Rabbit mAb #3133 40 µl W H M R 145 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Met (D1C2) XP® Rabbit mAb #8198 40 µl W IP IHC-P IHC-F IF-IC F H 140, 170 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

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.

Western Blotting

Western Blotting

Western blot analysis of cell extracts from HeLa cells, untreated or stimulated with HGF, using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb #3077 (upper) and Met (25H2) Mouse mAb #3127 (lower).

Western Blotting

Western Blotting

Western blot analysis of cell lysates of H4IIE cells, untreated or treated with HGF, using Phospho-Met (Tyr1349) (130H2) Rabbit mAb #3133 (upper), or Met (25H2) Mouse mAb #3127 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or HGF-treated, using Phospho-Met (Tyr1003) (13D11) Rabbit mAb #3135 (upper and middle) and Met (25H2) Mouse mAb #3127 (lower). The middle blot was treated with CIP phosphatase before antibody probing.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Gab1 Antibody #3232.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293 cells, untreated or treated with In1B for indicated times, using Phospho-Gab1 (Tyr307) Antibody #3234 (upper) or Gab1 Antibody #3232 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HT-29 (Met+), SK-BR-3 (Met-), and T-47D (Met-) cells using Met (D1C2) XP® Rabbit mAb #8198 (upper) or β-Actin Antibody #4967 (lower).


Description

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

Source / Purification

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.

Background

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.

Application References

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Protocols

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and/or U.S.S.N. 11/476,277) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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