Revision 1

#30590Store at -20C

1 个试剂盒

(4 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
PTEN (D4.3) XP® Rabbit mAb 9188 20 µl 54 kDa Rabbit IgG
Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb 9549 20 µl 54 kDa Rabbit IgG
PDK1 (D37A7) Rabbit mAb 5662 20 µl 58-68 kDa Rabbit IgG
Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb 3438 20 µl 58 to 68 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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

Description

The PTEN and PDK1 Antibody Sampler Kit II provides an economical means to evaluate two key enzymes that regulate multiple signaling pathways. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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 antibodies.

Background

PTEN (phosphatase and tensin homologue deleted on chromosome ten), also referred to as MMAC (mutated in multiple advanced cancers) phosphatase, is a tumor suppressor implicated in a wide variety of human cancers (1). PTEN encodes a 403 amino acid polypeptide originally described as a dual-specificity protein phosphatase (2). The main substrates of PTEN are inositol phospholipids generated by the activation of the phosphoinositide 3-kinase (PI3K) (3). PTEN is a major negative regulator of the PI3K/Akt signaling pathway (1,4,5). PTEN possesses a carboxy-terminal, noncatalytic regulatory domain with three phosphorylation sites (Ser380, Thr382, and Thr383) that regulate PTEN stability and may affect its biological activity (6,7). PTEN regulates p53 protein levels and activity (8) and is involved in G protein-coupled signaling during chemotaxis (9,10).
Phosphoinositide-dependent protein kinase 1 (PDK1) plays a central role in many signal transduction pathways (11,12), including the activation of Akt and the PKC isoenzymes p70 S6 kinase and RSK (13). Through its effects on these kinases, PDK1 is involved in the regulation of a wide variety of processes, including cell proliferation, differentiation, and apoptosis.

  1. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
  2. Myers, M.P. et al. (1997) Proc Natl Acad Sci USA 94, 9052-7.
  3. Myers, M.P. et al. (1998) Proc Natl Acad Sci USA 95, 13513-8.
  4. Wan, X. and Helman, L.J. (2003) Oncogene 22, 8205-11.
  5. Wu, X. et al. (1998) Proc Natl Acad Sci USA 95, 15587-91.
  6. Vazquez, F. et al. (2000) Mol Cell Biol 20, 5010-8.
  7. Torres, J. and Pulido, R. (2001) J Biol Chem 276, 993-8.
  8. Freeman, D.J. et al. (2003) Cancer Cell 3, 117-30.
  9. Funamoto, S. et al. (2002) Cell 109, 611-23.
  10. Iijima, M. and Devreotes, P. (2002) Cell 109, 599-610.
  11. Belham, C. et al. (1999) Curr Biol 9, R93-6.
  12. Toker, A. and Newton, A.C. (2000) Cell 103, 185-8.
  13. Williams, M.R. et al. (2000) Curr Biol 10, 439-48.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

    除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

    专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

    Revision 1
    #30590

    PTEN and PDK1 Antibody Sampler Kit II

    PTEN and PDK1 Antibody Sampler Kit II: Image 1 Expand Image
    使用 PTEN (D4.3) XP® Rabbit mAb #9188 对 HeLa 细胞的裂解物 (1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
    PTEN and PDK1 Antibody Sampler Kit II: Image 2 Expand Image
    使用 Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb (上图)和 Akt (pan) (C67E7) Rabbit mAb #4691(下图)对 PC3 细胞、HCT116 野生型和 HCT116 PDK1 -/- 细胞的提取物进行蛋白质印迹分析。(HCT116 野生型和 HCT116 PDK1 -/- 细胞由美国马里兰州巴尔的摩约翰霍普金斯大学的 Bert Vogelstein 博士惠赠)。
    PTEN and PDK1 Antibody Sampler Kit II: Image 3 Expand Image
    使用 PDK1 (D37A7) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
    PTEN and PDK1 Antibody Sampler Kit II: Image 4 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    PTEN and PDK1 Antibody Sampler Kit II: Image 5 Expand Image
    使用 PTEN (D4.3) XP® Rabbit mAb(上图)和 Akt (pan) (C67E7) Rabbit mAb #4691(下图),对不同细胞系的提取物进行蛋白质印迹分析。
    PTEN and PDK1 Antibody Sampler Kit II: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
    PTEN and PDK1 Antibody Sampler Kit II: Image 7 Expand Image
    使用 Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb 对 HeLa 和 NIH/3T3 细胞的提取物进行蛋白质印迹分析。在蛋白质印迹转移后,膜用 (-) 或不用 (+) 牛小肠磷酸酶 (CIP) 处理,以确定该抗体的磷酸特异性。
    PTEN and PDK1 Antibody Sampler Kit II: Image 8 Expand Image
    使用 Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb (上图)、PDK1 Antibody #3062(中间)或 Akt Antibody #9272(下图)对未经处理或已经 λ-磷酸酶处理的 PC3 细胞的提取物进行蛋白质印迹分析。
    PTEN and PDK1 Antibody Sampler Kit II: Image 9 Expand Image
    使用 PDK1 (D37A7) Rabbit mAb (上图)和 Akt (pan) (C67E7) Rabbit mAb #4691(下图)对 HCT 116 野生型和 HCT 116 PDK1-/- 细胞的提取物进行蛋白质印迹分析。(HCT 116 野生型和 HCT 116 PDK1-/- 细胞由美国马里兰州巴尔的摩约翰霍普金斯大学的 Bert Vogelstein 博士惠赠)。
    PTEN and PDK1 Antibody Sampler Kit II: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
    PTEN and PDK1 Antibody Sampler Kit II: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
    PTEN and PDK1 Antibody Sampler Kit II: Image 12 Expand Image
    使用 PTEN (D4.3) XP® Rabbit mAb 对石蜡包埋的人结肠细胞进行免疫组织化学分析。
    PTEN and PDK1 Antibody Sampler Kit II: Image 13 Expand Image
    使用 PTEN (D4.3) XP® Rabbit mAb 对 SignalSlide(TM) PTEN IHC Controls #8106(石蜡包埋的 LNCaP(左图)和 NIH/3T3(右图)细胞)进行免疫组织化学分析。