Revision 1

#9935Store at -20C

1 个试剂盒

(5 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
Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb 6943 20 µl 60 kDa Rabbit IgG
Non-phospho-Src (Tyr527) Antibody 2107 20 µl 60 kDa Rabbit 
Non-phospho-Src (Tyr416) (7G9) Mouse mAb 2102 20 µl 60 kDa Mouse IgG2b
Phospho-Src (Tyr527) Antibody 2105 20 µl 60 kDa Rabbit 
Src (32G6) Rabbit mAb 2123 20 µl 60 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

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

Description

The Src Antibody Sampler kit provides an economical means of evaluating total Src protein levels and its phosphorylation status. The kit contains enough primary and secondary antibodies to perform two Western blot experiments per 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 antibody.

Background

The Src family of protein tyrosine kinases, which includes Src, Lyn, Fyn, Yes, Lck, Blk, and Hck, are important in the regulation of growth and differentiation of eukaryotic cells (1). Src activity is regulated by tyrosine phosphorylation at two sites, but with opposing effects. While phosphorylation at Tyr416 in the activation loop of the kinase domain upregulates enzyme activity, phosphorylation at Tyr527 in the carboxy-terminal tail by Csk renders the enzyme less active (2).
Lyn is a member of the Src family that is predominantly expressed in hematopoietic cells (3). Lyn participates in signaling from multiple cell surface receptors such as the B cell Ag receptor (BCR) and CD40 (4).
Lck is essential for T-lymphocyte activation and differentiation (5,6). The activity of Lck is regulated by protein kinases and phosphatases. Phosphorylation of the C-terminal tyrosine 505 serves to downregulate Lck catalytic activity, while phosphorylation at tyrosine 394 leads to an increase in Lck activity (7).

  1. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
  2. Hunter, T. (1987) Cell 49, 1-4.
  3. Yamanashi, Y. et al. (1989) Proc Natl Acad Sci U S A 86, 6538-42.
  4. Yamanashi, Y. et al. (1991) Science 251, 192-4.
  5. Molina, T.J. et al. (1992) Nature 357, 161-4.
  6. Straus, D.B. and Weiss, A. (1992) Cell 70, 585-93.
  7. Chow, L.M. et al. (1993) Nature 365, 156-60.

Background References

    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.

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    Revision 1
    #9935

    Src Antibody Sampler Kit

    Src Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from HepG2 cells using Non-phospho-Src (Tyr416) (7G9) Mouse mAb #2102. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:50 和 1:250 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:50(蓝线)和 1:250(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。
    Src Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from 3T3 cells using Phospho-Src (Tyr527) Antibody #2105. 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。
    Src Antibody Sampler Kit: Image 3 Expand Image
    使用 Non-phospho-Src (Tyr416) (7G9 ) Mouse mAb(上)、v-Src antibody(中)或 Phospho-Src (Tyr416) Antibody #2101(下),对过钒酸钠处理(1 mM,5 分钟)且经 Src 的结构活性形式(调节性酪氨酸 527 残基已突变为苯基丙氨酸)稳定转染的 NIH/3T3 细胞、A431 细胞和 HepG2 细胞提取物进行蛋白质印迹分析。
    Src Antibody Sampler Kit: Image 4 Expand Image
    使用 Phospho-Src (Tyr527) Antibody (A,B) 或 v-Src antibody (C,D) ,对 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,使用牛小肠碱性磷酸酶 (CIP) (B,D) 处理膜来确认该抗体的磷酸特异性。
    Src Antibody Sampler Kit: Image 5 Expand Image
    使用 Non-phospho-Src (Tyr527) Antibody(上)或 v-Src antibody(下)对未处理或钒酸钠处理(1 mM,15 分钟)的 A431 细胞提取物进行蛋白质印迹分析。
    Src Antibody Sampler Kit: Image 6 Expand Image
    使用 Src (32G6) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
    Src Antibody Sampler Kit: Image 7 Expand Image
    使用 Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb(上)或 Src (36D10) Rabbit mAb #2109(下),对血清饥饿或经人 Platelet-Derived Growth Factor BB hPDGF-BB #8912 处理(100 ng/ml,15 分钟)的 NIH/3T3 细胞提取物进行蛋白质印迹分析。
    Src Antibody Sampler Kit: Image 8 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。
    Src Antibody Sampler Kit: Image 9 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*,它可在酶催化的分解期间发光。
    Src Antibody Sampler Kit: Image 10 Expand Image
    使用 Phospho-Src (Tyr527) Antibody 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析,显示细胞膜和细胞浆定位。