Cell Signaling Technology

Product Pathways - Motif Antibodies

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

Applications Reactivity Source Isotype
W IP IHC-P IF-F IF-IC IF-P F E-P All Mouse IgG1

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

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)

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)

IHC-P (paraffin)

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


IHC-P (paraffin)

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 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

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

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

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

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

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Companion Products

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.

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