Revision 1

#12718Store 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
Calnexin (C5C9) Rabbit mAb 2679 20 µl 90 kDa Rabbit IgG
ERp72 (D70D12) XP® Rabbit mAb 5033 20 µl 72 kDa Rabbit IgG
PDI (C81H6) Rabbit mAb 3501 20 µl 57 kDa Rabbit 
RCAS1 (D2B6N) XP® Rabbit mAb 12290 20 µl 32 kDa Rabbit IgG
Syntaxin 6 (C34B2) Rabbit mAb 2869 20 µl 32 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 ER and Golgi-Associated Marker Proteins Antibody Sampler Kit contains reagents to examine proteins that help regulate protein folding and vesicle trafficking. This kit includes enough antibody 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 antibody.

Background

Secretory and transmembrane proteins are synthesized on polysomes and translocate into the endoplasmic reticulum (ER) where they are often modified by the formation of disulfide bonds, amino-linked glycosylation, and folding. The ER contains a pool of molecular chaperones to help proteins fold properly. Calnexin is a calcium-binding, ER membrane protein that ensures proper protein folding by retaining newly synthesized glycoproteins within the ER l (1-3). The specificity of calnexin for a subset of glycoproteins is defined by a lectin site, which binds an early oligosaccharide intermediate on the folding glycoprotein (3). Many secretory proteins require the formation of intra- or inter-molecular disulfide bonds to reach their native conformation (4). Protein disulfide isomerase (PDI) catalyzes the formation and isomerization of disulfide bonds during oxidative protein folding (5). The ER-protein Ero1 oxidizes PDI through disulfide exchange, which is followed by PDI-catalyzed disulfide bond formation in folding proteins (6). The ER stress protein 72 (ERp72) contains three thioredoxin homology domains and plays a role in the formation and isomerization of disulfide bonds (7,8).

The tumor-associated antigen RCAS1 negatively regulates cytotoxic T lymphocyte (CTL) cytolytic activity, which impacts vesicle formation, secretion, and protein glycosylation (9-12). Overexpression of RCAS1 impairs CTL cytolytic function by negatively regulating trans-Golgi to secretory lysosome protein trafficking, leading to a delay in ER to Golgi vesicle transport and mislocalization of ER quality control and glycosylation proteins. As a result, RCAS1 induces deposition of tumor-associated glycan antigens on the cell surface, which may contribute to tumor pathogenesis through the mediation of adhesion, invasion, and metastasis (13,14). Syntaxin 6 is a ubiquitously expressed S25C family member of the SNARE proteins (15,16) that is localized to the trans-Golgi and within endosomes. It regulates membrane trafficking by partnering with a variety of other SNARE proteins (17-19) and is involved in the regulation of GLUT4 trafficking, neutrophil exocytosis, and granule secretion (20-22).

  1. Rajagopalan, S. et al. (1994) Science 263, 387-90.
  2. Bergeron, J.J. et al. (1994) Trends Biochem Sci 19, 124-8.
  3. Williams, D.B. (2006) J Cell Sci 119, 615-23.
  4. Huppa, J.B. and Ploegh, H.L. (1998) Cell 92, 145-8.
  5. Ellgaard, L. and Ruddock, L.W. (2005) EMBO Rep 6, 28-32.
  6. Tu, B.P. and Weissman, J.S. (2004) J Cell Biol 164, 341-6.
  7. Mazzarella, RA et al. (1990) J Biol Chem 265(2), 1094-101.
  8. Satoh, M et al. (2005) Cell Stress Chaperones 10(4), 278-84
  9. Rüder, C. et al. (2009) J Clin Invest 119, 2184-203.
  10. Reimer, T.A. et al. (2005) BMC Cancer 5, 47.
  11. Wolf, J. et al. (2010) FASEB J 24, 4000-19.
  12. Engelsberg, A. et al. (2003) J Biol Chem 278, 22998-3007.
  13. Bock, J.B. et al. (2001) Nature 409, 839-41.
  14. Bock, J.B. et al. (1996) J Biol Chem 271, 17961-5.
  15. Wendler, F. and Tooze, S. (2001) Traffic 2, 606-11.
  16. Bock, J.B. et al. (1997) Mol Biol Cell 8, 1261-71.
  17. Mallard, F. et al. (2002) J Cell Biol 156, 653-64.
  18. Perera, H.K. et al. (2003) Mol Biol Cell 14, 2946-58.
  19. Shewan, A.M. et al. (2003) Mol Biol Cell 14, 973-86.
  20. Martín-Martín, B. et al. (2000) Blood 96, 2574-83.
  21. Wendler, F. et al. (2001) Mol Biol Cell 12, 1699-709.
  22. Kuliawat, R. et al. (2004) Mol Biol Cell 15, 1690-701.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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.

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    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

    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。
    Revision 1
    #12718

    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit

    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 1 Expand Image
    使用 Calnexin (C5C9) Rabbit mAb #2679 对来自 HepG2 细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像（左图）显示一抗稀释比例在 1:50 和 1:250 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 1:50（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。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 2 Expand Image
    以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（虚线）作为对照，使用 ERp72 (D70D12) XP® Rabbit mAb（实线）对 Hep G2 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 3 Expand Image
    使用 RCAS1 (D2B6N) XP® Rabbit mAb，对不同细胞系的提取物进行蛋白质印迹分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 4 Expand Image
    使用 Calnexin (C5C9) Rabbit mAb 对 PANC1、HepG2 和 A204 细胞提取物进行蛋白质印迹法分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 5 Expand Image
    使用 Syntaxin 6 (C34B2) Rabbit mAb，对不同细胞系的提取物进行蛋白质印迹分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 6 Expand Image
    使用 PDI (C81H6) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 7 Expand Image
    使用 ERp72 (D70D12) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    ER and Golgi-Associated Marker Proteins 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。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 9 Expand Image
    使用 RCAS1 (D2B6N)XP® Rabbit mAb，对转染空载（-）或转染带有 Myc/DDK 标签的全长人 RCAS1（hRCAS1-Myc/DDK；+）表达载体的 293T 细胞的提取物进行蛋白质印迹分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 10 Expand Image
    在对照肽（左）或抗原特异性肽（右）存在的情况下，使用 Calnexin (C5C9) Rabbit mAb 对石蜡包埋的人乳腺癌组织进行免疫组织化学分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 11 Expand Image
    使用 Syntaxin 6 (C34B2) Rabbit mAb（绿色），对 MCF7 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记（红色）。蓝色伪彩 = DRAQ5™ （DNA 荧光染料）。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 12 Expand Image
    使用 PDI (C81H6) Rabbit mAb 对石蜡包埋的人肺癌进行免疫组织化学分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 13 Expand Image
    使用 ERp72 (D70D12) XP® Rabbit mAb（绿色）和 β-Actin (8H10D10) Mouse mAb #3700（红色）对 PANC-1 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 14 Expand Image
    使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（泳道 2）或 RCAS1 (D2B6N) XP® Rabbit mAb（泳道 3），对 HCT 116 提取物 RCAS1 进行免疫沉淀。泳道 1 是 10% 输入对照。使用 RCAS1 (D2B6N) XP® Rabbit mAb 进行蛋白质印迹分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 15 Expand Image
    使用 Calnexin (C5C9) Rabbit mAb（绿色）对 HeLa 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝已用 DY-554 phalloidin（红色）标记。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 16 Expand Image
    使用 PDI (C81H6) Rabbit mAb 对石蜡包埋的人淋巴癌进行免疫组织化学分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 17 Expand Image
    使用 RCAS1 (D2B6N) XP® Rabbit mAb（绿色），对未经处理（左）或经 Brefeldin A #9972 处理（5 μg/ml，1 小时；右）的 MCF7 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin（红色）标记肌动蛋白纤丝。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 18 Expand Image
    使用 PDI (C81H6) Rabbit mAb 对石蜡包埋的小鼠脾脏进行免疫组织化学分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 19 Expand Image
    使用浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（红色）作为 RCAS1 (D2B6N) XP® Rabbit mAb（蓝色）的对照，对 Jurkat 细胞进行流式细胞分析。
    ER and Golgi-Associated Marker Proteins Antibody Sampler Kit: Image 20 Expand Image
    使用 PDI (C81H6) Rabbit mAb（绿色）和 β-Actin (8H10D10) Mouse mAb #3700（红色）对 NIH/3T3 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。