| Application |
Dilution |
Species-Reactivity |
Sensitivity |
Isotype |
| W |
|
Human, Mouse, Rat |
Endogenous |
Rabbit |
| IP |
|
| IHC-P |
|
| IHC-F |
|
| IF-IC |
|
| F |
|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Specificity / Sensitivity
Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb detects endogenous levels of Met only when phosphorylated at Tyr1234/1235. This antibody may cross-react with overexpressed tyrosine phosphorylated Src proteins in Western blot. The use of this antibody for IF and F applications are only recommended for cells over expressing phospho-Met (Y1234/1235).
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1234/1235 of human Met.
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 (upper) and Met (25H2) Mouse mAb #3127 (lower).
Western Blotting
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.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.
IHC-P (paraffin)
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.
IHC-P (paraffin)
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.
IHC-P (paraffin)
Immunohisochemical analysis of paraffin-embedded HCC827 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.
IHC-P (paraffin)
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)].
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded papillary renal cell carcinoma using Phospho-Met (Tyr1234/1225) (D26) XP® Rabbit mAb.
IHC-F (frozen)
Immunohistochemical analysis of frozen MKN45 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.
IF-IC
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).
Flow Cytometry
Flow cytometric analysis of MKN-45 cells, untreated (green) or treated with SU11274 (blue).
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. The addition of a phosphate at cytoplasmic Tyr1003 is essential for Met protein ubiquitination and degradation (4). Phosphorylation at Tyr1234/1235 in the Met kinase domain is critical for kinase activation. Phosphorylation at Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Research studies have shown that altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon, and breast. Thus, investigators have concluded that Met is an attractive potential cancer therapeutic and diagnostic target (6,7).
- Cooper, C.S. et al. (1984) Nature 311, 29-33.
- Bottaro, D.P. et al. (1991) Science 251, 802-4.
- Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
- Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
- Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
- Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
- Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.
Application References
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoSitePlus® protein, site, and accession data:
