Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-F, IF-IC, FC-FP, E-P

REACTIVITY:

All

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Mouse IgG1

Product Information

Product Usage Information

The phosphorylated form of the peptide can be detected with Phospho-Tyrosine mAb (P-Tyr-100) #9411. Sample kinase protocol is attached.

Application Dilution
Western Blotting 1:2000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:2400 - 1:9600
Immunofluorescence (Frozen) 1:1600 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:1600 - 1:6400
Peptide ELISA (DELFIA) 1:4000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #80379.

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.

Species Reactivity:

All Species Expected

Source / Purification

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

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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) E-P: Peptide ELISA (DELFIA)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

Revision 6
#9411

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

Western Blotting Image 1: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
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.
No image available
Immunohistochemistry Image 1: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 2: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 3: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft untreated (left) or lambda-phosphatase-treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 4: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 5: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 6: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis using Phospho-Tyrosine Mouse mAb (P-Tyr-100) on SignalSlide Phospho-EGF Receptor IHC Controls #8102 (paraffin-embedded KYSE450 cell pellets, untreated (left) or EGF-treated (right)).
Immunohistochemistry Image 7: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded human soft tissue squamous cell carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 8: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded normal human kidney using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 9: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded normal human lung using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunohistochemistry Image 10: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
Immunofluorescence Image 1: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
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.)
Immunofluorescence Image 2: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
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).
Flow Cytometry Image 1: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
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).
Product Image 1: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Expand Image
Phospho-Tyrosine Mouse mAb (P-Tyr-100) (HRP Conjugate) ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (Y* denotes phosphorylated tyrosine.)