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REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous 145 Mouse IgG1
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Flow cytometric analysis of live Ramos cells (red) and live MKN-45 cells (blue) using Met (11C4) Mouse mAb (Flow Specific).

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Flow Cytometry

A. Solutions and Reagents

B. Immunostaining

NOTE: Account for isotype matched controls for monoclonal antibodies or species matched IgG for polyclonal antibodies. Count cells using a hemacytometer or alternative method.

  1. Collect cells by centrifugation and aspirate supernatant.
  2. Aliquot 0.5–1x106 cells into each assay tube (by volume).
  3. Add 2–3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl Incubation Buffer per assay tube.
  5. Block in Incubation Buffer for 10 minutes on ice.
  6. Add the unconjugated primary antibody at the appropriate dilution to the assay tubes (see individual antibody data sheet for the dilution recommendations).
  7. Incubate for 1 hour on ice.
  8. Rinse as before in Incubation Buffer by centrifugation.
  9. Resuspend cells in fluorochrome-conjugated secondary antibody* diluted in total volume Incubation Buffer at the recommended dilution.
  10. Incubate for 30 minutes on ice.
  11. Rinse as before in Incubation Buffer by centrifugation.
  12. Resuspend cells in 0.5 ml PBS and analyze on flow cytometer.

posted Februay 2011

protocol id: 58

Product Usage Information

Note: This antibody can only be used to detect live cells.


Application Dilutions
Flow Cytometry 1:50

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.

Specificity / Sensitivity

Met (11C4) Mouse mAb (Flow Specific) detects endogenous levels of total Met protein.


Species Reactivity: Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with human cancer cell lines.

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).


1.  Cooper, C.S. et al. (1984) Nature 311, 29-33.

2.  Bottaro, D.P. et al. (1991) Science 251, 802-4.

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.  Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.

7.  Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.


Entrez-Gene Id 4233
Swiss-Prot Acc. P08581


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