Cell Signaling Technology

Product Pathways - Screening Technologies

Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb #8954

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F All Endogenous N/A Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb recognizes a broad range of tyrosine-phosphorylated proteins and peptides. This antibody does not cross-react with proteins or peptides containing phospho-Ser or phospho-Thr residues.

Source / Purification

Monoclonal antibody is produced by immunizing animals with phospho-tyrosine containing peptides.

Western Blotting

Western Blotting

Western blot analysis of extracts from A-431 cells, untreated (-) or treated with Human Epidermal Growth Factor (hEGF) #8916 (100 ng/ml, 5 min; +), using Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb. Western blot image was obtained using the Odyssey® Infrared Imaging System (LI-COR® Biotechnology).

IP

IP

Immunoprecipitation of phospho-tyrosine proteins from A-431 cell extracts, untreated (-) or treated with Human Epidermal Growth Factor (hEGF) #8916 (100 ng/ml, 5 min; +) (lanes 3 and 4), using Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb. Western blot analysis was performed using the same antibody. Lanes 1 and 2 are 10% input.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K-562 cells, untreated (green) or Gleevec®-treated (blue), using Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, serum-starved (left), treated with H2O2 (2 mM, 10 min; middle), or treated with H2O2 followed by λ phosphatase (right), using Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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.

  1. Schlessinger, J. (2000) Cell 103, 211-25.
  2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
  3. Ward, S.G. et al. (1992) J Biol Chem 267, 23862-9.
  4. Glenney, J.R. et al. (1988) J Immunol Methods 109, 277-85.

Application References

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Companion Products

This product is protected by 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) owned by Cell Signaling Technology.

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

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