Product Pathways - Motif Antibodies
Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411
Looking for in-depth proteomic analysis of regulatory PTMs? PTMScan® Services are now available.
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| W IP IHC-P IF-P IF-F IF-IC F E-P | All | Endogenous | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-P=Immunofluorescence (Paraffin)
IF-F=Immunofluorescence (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.
Protocols
- 9411:
- ELISA Peptide, Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting
Specificity / Sensitivity
Phospho-Tyrosine Mouse mAb (P-Tyr-100) is a high affinity antibody. ELISAs against a wide variety of phosphopeptides indicate that 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 .
Western Blotting
Western blot analysis of extracts from Jurkat cells treated with 1 mM pervanadate for 30 minutes prior to lysis, using P-Tyr-100 Phospho-Tyrosine Mouse mAb. Proteins were separated by 2-D electrophoresis prior to blotting.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma 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 to a nonspecific negative control antibody (red).
IF-IC
Immunofluorescent analysis of Swiss NIH/3T3 cells, serum-starved and stimulated with lysophosphatidic acid (LPA) (10 µM for 10 minutes) and fixed with PFA, using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red) and phalloidin for F-actin (green) 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 analysis of HeLa cells either stimulated with 20% serum (left) or untreated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Actin filaments have been labeled with fluorescein phalloidin. Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
IF-P
Confocal immunofluorescent analysis of paraffin-embedded human lung adenocarcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
ELISA-Peptide
Phospho-Tyrosine Mouse mAb (P-Tyr-100) (HRP Conjugate) ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (Y* denotes phosphorylated tyrosine.)
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
- Smith, J.A. et al. (2005) Cancer Res 65, 1027-34. Applications: Western Blotting
- Ye, Z.J. et al. (2005) Proc Natl Acad Sci U S A 102, 18461-6. Applications: Western Blotting
- Sharma, A. et al. (2003) Mol Cell Proteomics 2, 1217-24. Applications: IP Western Blotting
- Li, Y. et al. (2003) Mol Cell Biol 23, 6255-66. Applications: IP Western Blotting
- Kogata, N. et al. (2003) Mol Biol Cell 14, 3553-64. Applications: Western Blotting
- del Rincón, S.V. et al. (2003) Oncogene 22, 3353-60. Applications: Western Blotting
- Goldberg, G.S. et al. (2003) J Biol Chem 278, 46533-40. Applications: Western Blotting
- Klein, R.D. and Fischer, S.M. (2002) Carcinogenesis 23, 217-21. Applications: Western Blotting
- Perez, O.D. and Nolan, G.P. (2002) Nat Biotechnol 20, 155-62. Applications: Flow Cytometry
- Kaluz, S. et al. (2002) Cancer Res 62, 4469-77. Applications: IP
- Kato, R. et al. (2002) Clin Cancer Res 8, 2455-62. Applications: IP
- Asao, H. et al. (2001) J Immunol 167, 1-5. Applications: Western Blot
- Stewart, D.M. et al. (2001) Endocrinology 142, 98-107. Applications: Western Blotting
- Farhang-Fallah, J. et al. (2000) J Biol Chem 275, 40492-7. Applications: Western Blot
- Chang, C.I. et al. (2000) J Immunol 165, 2134-41. 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
- 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)
- 9419 Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Sepharose Bead Conjugate)
- 9386 Phospho-Threonine (42H4) Mouse mAb
- 9614 Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb
- 9624 Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb
- 8102 SignalSlide® Phospho-EGF Receptor IHC Controls
- 8112 SignalStain® Antibody Diluent
- 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
- 8125 SignalStain® Boost IHC Detection Reagent (HRP, Mouse)
- 5425 Normal Goat Serum
- 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
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.