Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate)
Product information| Code | Name | Size | Quantity | Price | |
|---|---|---|---|---|---|
9415S |
Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) |
100 µl (50 tests) | - | Unavailable in your region |
Product Pathways - Motif Antibodies
Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) #9415
PhosphoSitePlus® protein, site, and accession data:
| No. | Size | Price | ||
|---|---|---|---|---|
| 9415S | 100 µl (50 tests) | --- | In Stock | --- |
| 9415 | carrier free and custom formulation / quantity | email request | ||
| Application | Dilution | Species-Reactivity | Sensitivity | Isotype |
|---|---|---|---|---|
| IF-IC | 1:50 | All Species Expected | Endogenous | Mouse IgG1 |
| F | 1:50 |
Species cross-reactivity is determined by western blot using the unconjugated antibody.
Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Protocols
- 9415:
- Immunofluorescence, Flow
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.
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).
Flow Cytometry
Flow cytometric analysis of K-562 cells, untreated (green) or treated with with Imatinib #9084 (10 uM, 2.5 hr; blue) using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) (solid lines) or concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 647 Conjugate) #4843 (dashed lines).
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.
Application References
- Kalland, M.E. et al. (2011) J Immunol 187, 5233-45. Applications: Flow Cytometry.
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Companion Products
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.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 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.
The supporting documents available for this product can be downloaded below.