Product Pathways - Motif Antibodies

Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411

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Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  IF-P=Immunofluorescence (Paraffin)  F=Flow Cytometry  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

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 derived from mice immunized with phospho-tyrosine-containing peptides (KLH-coupled).

Western Blotting

P-Tyr-100 Phospho-Tyrosine Mouse mAb Antibody: Western blot analysis of extracts from Jurkat cells treated with 1 mM pervanadate for 30 minutes prior to lysis. Proteins were separated by 2-D electrophoresis prior to blotting.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing staining of proteins with phosphorylated tyrosine residues, using Phospho-Tyrosine Mouse mAb (P-Tyr-100).

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Tyrosine Mouse mAb (P-Tyr-100).

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft untreated (left) or lambda-phosphatase-treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).

Flow Cytometry

Flow cytometric analysis of K562 cells, untreated (green) or Gleevec®- treated (blue), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) compared with a nonspecific negative control antibody (red).

IF-IC

Immunofluorescent analysis of serum-starved Swiss NIH/3T3 cells, stimulated with lysophosphatidic acid (LPA) (10 µM for 10 minutes), fixed with PFA and stained with phalloidin for F-actin (green) and Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). LPA causes heavy tyrosine phosphorylation of proteins in focal adhesions, present at the tips of actin stress fibers. (Provided by Dr. Harry Mellor, University of Bristol.)

IF-IC

Confocal immunofluorescence images of HeLa cells 20% serum-treated (left) or untreated (right) and labeled with Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Actin filaments have been labeled with fluorescein phalloidin. Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

IF-F

Confocal immunofluorescence image of paraffin-embedded human lung adenocarcinoma labeled with Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

ELISA-Peptide

Phospho-Tyrosine Mouse mAb (P-Tyr-100) DELFIA® Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (Y* denotes phosphorylated tyrosine.) (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Background

Tyrosine phosphorylation plays a key role in cellular signaling (1). 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 Mouse mAbs developed by CST (P-Tyr-100, #9411 and P-Tyr-102, #9416) provide exceptionally sensitive new tools of increased utility for studying tyrosine phosphoyrlation and monitoring tyrosine kinase activity in high throughput drug discovery.

1. Schlessinger, J. (2000) Cell 103, 211-225.
2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-365.
3. Ward, S.G. et al. (1992) J. Biol. Chem. 267, 23862-23869.
4. Glenney, J.R. et al. (1988) J. Immunol. Methods. 109, 277-285.

Application References

• Chang, C. I. et al. (2000) The involvement of tyrosine kinases, cyclic AMP/protein kinase A, and p38 mitogen-activated protein kinase in IL-13-mediated arginase I induction in macrophages: its implications in IL-13-inhibited nitric oxide production. J. Immunol. 165, 2134-2141. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blot
• Farhang-Fallah, J. et al. (2000) Cloning and characterization of PHIP, a novel insulin receptor substrate-1 pleckstrin homology domain interacting protein. J. Biol. Chem. 275, 40492-40497. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blot
• Stewart, D. M. et al. (2001) Interferon-tau activates multiple signal transducer and activator of transcription proteins and has complex effects on interferon-responsive gene transcription in ovine endometrial epithelial cells. Endocrinology 142 (1), 98-107. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blotting
• Klein, R.D. and Fischer, S.M. (2002) Black tea polyphenols inhibit IGF-I-induced signaling through Akt in normal prostate epithelial cells and Du145 prostate carcinoma cells. Carcinogenesis 23 (1), 217-221. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blotting
• Perez, O.D. and Nolan, G.P. (2002) Simultaneous measurement of multiple active kinase states using polychromatic flow cytometry. Nat. Biotechnol. 20 (2), 155-162. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Flow Cytometry
• Kaluz, S. et al. (2002) Lowered oxygen tension induces expression of the hypoxia marker MN/carbonic anhydrase IX in the absence of hypoxia-inducible factor 1 alpha stabilization: a role for phosphatidylinositol 3'-kinase. Cancer Res. 62, 4469-4477. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: IP
• Li, Y. et al. (2003) Morphine induces desensitization of insulin receptor signaling. Mol. Cell. Biol. 23 (17), 6255-6266. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: IP Western Blotting
• Kogata, N. et al. (2003) Identification of Fer tyrosine kinase localized on microtubules as a platelet endothelial cell adhesion molecule-1 phosphorylating kinase in vascular endothelial cells. Mol. Biol. Cell 14, 3553-3564. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blotting
• del Rincon, S. V. et al. (2003) Retinoic acid-induced growth arrest of MCF-7 cells involves the selective regulation of the IRS-1/PI 3-kinase/AKT pathway. Oncogene 22, 3353-3360. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blotting
• Goldberg, G. S. et al. (2003) Src phosphorylates Cas on tyrosine 253 to promote migration of transformed cells. J. Biol. Chem. 278 (47), 46533-46540. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blotting
• Sharma, A. et al. (2003) Functional Interaction Trap: A strategy for validating the functional consequences of tyrosine phosphorylation of specific substrates in vivo. Mol. Cell. Proteomics 2 (11), 1217-1224. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: IP Western Blotting
• Kato, R. et al. (2002) A new type of antimetastatic peptide derived from fibronectin. Clin. Cancer Res. 8, 2455-2462. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: IP
• Asao, H. et al. (2001) Cutting edge: The common gamma-chain is an indispensable subunit of the IL-21 receptor complex. J. Immonol. 167, 1-5. This article references the use of Phospho-Tyrosine Mouse mAb (P-Tyr-100) in the following applications: Western Blot

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

• 7900 PhosphoScan® Kit (P-Tyr-100)
• 9414 Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 488 Conjugate)
• 9415 Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate)
• 9416 Phospho-Tyrosine Mouse mAb (P-Tyr-102)
• 9417 Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Biotinylated)
• 9418 Phospho-Tyrosine Mouse mAb (P-Tyr-102) (Fluorescein Conjugate)
• 9419 Immobilized Phospho-Tyrosine Mouse mAb (P-Tyr-100)
• 9386 Phospho-Threonine (42H4) Mouse mAb
• 9614 Phospho-Akt Substrate (RXRXXS/T) (110B7E) Rabbit mAb
• 9624 Phospho-PKA Substrate (RRXS/T) (100G7E) Rabbit mAb
• 7902 Phospho-Tyrosine Mouse mAb (P-Tyr-100) Beads(for use with PhosphoScan® Technology)
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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 commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.