Revision 1
Cell Signaling Technology

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Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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

#P08581

Entrez-Gene Id:

4233

Product Includes Quantity Reactivity MW(kDa) Isotype
Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb 3077 100 µl H M R 145 Rabbit 
Met (D1C2) XP® Rabbit mAb 8198 100 µl H 140, 170 Rabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

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.

Background

Met, a high affinity tyrosine kinase receptor for hepatocyte growth factor (HGF, also known as scatter factor) is a disulfide-linked heterodimer made of 45 kDa α- and 145 kDa β-subunits (1,2). The α-subunit and the amino-terminal region of the β-subunit form the extracellular domain. The remainder of the β-chain spans the plasma membrane and contains a cytoplasmic region with tyrosine kinase activity. Interaction of Met with HGF results in autophosphorylation at multiple tyrosines, which recruit several downstream signaling components, including Gab1, c-Cbl, and PI3 kinase (3). These fundamental events are important for all of the biological functions involving Met kinase activity. The addition of a phosphate at cytoplasmic Tyr1003 is essential for Met protein ubiquitination and degradation (4). Phosphorylation at Tyr1234/1235 in the Met kinase domain is critical for kinase activation. Phosphorylation at Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Research studies have shown that altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon, and breast. Thus, investigators have concluded that Met is an attractive potential cancer therapeutic and diagnostic target (6,7).

  1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
  2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
  3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
  4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
  5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
  6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
  7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus is a registered 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 1
    #8218

    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet

    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from HT-29 untreated cells using Met (D1C2) XP® Rabbit mAb #8198 lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 2 Expand Image
    Western blot analysis of cell extracts from HeLa cells, untreated or stimulated with HGF, using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (upper) and Met (25H2) Mouse mAb #3127 (lower).
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 3 Expand Image
    Western blot analysis of extracts from HT-29 (Met+), SK-BR-3 (Met-), and T-47D (Met-) cells using Met (D1C2) XP® Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 4 Expand Image
    Western blot analysis of purified active Ron kinase using a Phospho-Ron (Ser1394) Antibody (A), a Phospho-Ron (Tyr1238/1239) Antibody (B), Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (C) and Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (D). This demonstrates that Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb does not cross-react with phospho-Ron by western analysis.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 5 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1) or Met knockout HeLa cells (lane 2) using Met (D1C2) XP® Rabbit mAb #8198. The absence of signal in the Met knockout HeLa cells confirms specificity of the antibody for Met.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 7 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Met (D1C2) XP® Rabbit mAb performed on the Leica® Bond Rx.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded xenografts from 3T3-Met (left) and 3T3-Ron cells (right) using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb, indicating that this antibody does not cross-react with activated Ron by immunohistochemistry. Image courtesy of Pfizer, Inc.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Met (D1C2) XP® Rabbit mAb performed on the Leica® Bond Rx.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 10 Expand Image
    Immunohistochemical analysis on Src-transfected NIH/3T3 cells, using a Phospho-Src Family (Tyr416) Antibody (left) or Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (right), indicating that the antibody does not cross-react with Src phosphorylated at Tyr416 via immunohistochemistry.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human metastatic lung carcinoma using Met (D1C2) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 12 Expand Image
    Immunohisochemical analysis of paraffin-embedded HCC827 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Met (D1C2) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 14 Expand Image
    Immunohistochemical analysis using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb on SignalSlide Phospho-Met (1234/1235) IHC Controls #8118 [MKN45 cells, untreated (left) or SU11274-treated (right)].
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded human papillary renal cell carcinoma using Met (D1C2) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded papillary renal cell carcinoma using Phospho-Met (Tyr1234/1225) (D26) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded cell pellets, MKN-45 (left) and T-47D (right), using Met (D1C2) XP® Rabbit mAb.
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 18 Expand Image
    Confocal immunofluorescence analysis of MKN45 cells, untreated (left) or treated with SU11274 (1 μM, 3 hours; right), using Phospho-MET (Tyr1234/Tyr1235) (D26) XP® Rabbit mAb. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 19 Expand Image
    Confocal immunofluorescent analysis of HT-29 and T-47D cells using Met (D1C2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 20 Expand Image
    Flow cytometric analysis of MKN-45 cells, untreated (green) or treated with SU11274 (blue).
    PhosphoPlus® Met (Tyr1234/Tyr1235) Antibody Duet: Image 21 Expand Image
    Flow cytometric analysis of fixed and permeabilized Ramos cells (blue, negative) and MKN-45 cells (green, positive) using Met (D1C2) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.