Product Pathways - Motif Antibodies
Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) #9415
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| IF-IC F | All | Endogenous | Mouse IgG1 |
Applications Key:
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.
Protocols
- 9415:
- Flow, Immunofluorescence
Specificity / Sensitivity
Phospho-Tyrosine Mouse mAb (P-Tyr-100) is a high affinity antibody. ELISAs against a wide variety of phosphopeptides indicate that (a) P-Tyr-100 binds phospho-Tyr in a manner largely independent of the surrounding amino acid sequence. 2D gel Western blot analysis of pervanadate-treated cell extracts also shows that P-Tyr-100 interacts with a broad range of tyrosine-phosphorylated proteins. P-Tyr-100 does not cross-react with peptides containing phospho-Ser or phospho-Thr. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)
Source / Purification
Monoclonal antibody is produced by immunizing animals with phospho-tyrosine-containing peptides . The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6.
Flow Cytometry
Flow cytometric analysis of K562 cells, untreated (green) or Gleevec®- treated (blue), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) compared with a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent image of two Gefinitib (Iressa) treated non-small cell lung cancer cell lines. HCC827 cells have the E746_A750 deletion in exon 19 of the EGFR gene, and are highly sensitive to Gefitinib. H1975 cells have the (T790M) mutation that confers Gefitinib-resistance. Both cell lines were treated and then double-labeled with Phospho-S6 Ribosomal Protein Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854 and Phospho-Tyrosine Mouse mAb (Alexa Fluor® 647 Conjugate) #9415. Untreated HCC827 (A) and H1975 (C) cells show bright phospho-S6 (green) and phospho-tyrosine (red pseudocolor) label. Phospho-S6 and phospho-tyrosine signals dramatically decrease following Gefitinib treatment in HCC827 cells (B), with little or no change in the Gefitinib-resistant H1975 cells (D).
Description
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411.
Background
Tyrosine phosphorylation plays a key role in cellular signaling (1). Research studies have shown that in cancer, unregulated tyrosine kinase activity can drive malignancy and tumor formation by generating inappropriate proliferation and survival signals (2). Antibodies specific for phospho-tyrosine (3,4) have been invaluable reagents in these studies. The phospho-tyrosine monoclonal antibodies developed by Cell Signaling Technology are exceptionally sensitive tools for studying tyrosine phosphorylation and monitoring tyrosine kinase activity in high throughput drug discovery.
- Schlessinger, J. (2000) Cell 103, 211-25.
- Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
- Ward, S.G. et al. (1992) J Biol Chem 267, 23862-9.
- Glenney, J.R. et al. (1988) J Immunol Methods 109, 277-85.
Application References
- Kalland, M.E. et al. (2011) J Immunol 187, 5233-45. Applications: Flow Cytometry
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9414 Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 488 Conjugate)
- 9411 Phospho-Tyrosine Mouse mAb (P-Tyr-100)
- 9416 Phospho-Tyrosine Mouse mAb (P-Tyr-102)
- 9417 Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Biotinylated)
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.
For Research Use Only. Not For Use In Diagnostic Procedures.
