Revision 4

#29603Store at -20C

1 个试剂盒

(7 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
Fas (C18C12) Rabbit mAb 4233 20 µl 40-50 kDa Rabbit IgG
TNF-R1 (C25C1) Rabbit mAb 3736 20 µl 55 kDa Rabbit IgG
TNF-R2 (E8D7P) Rabbit mAb 72337 20 µl 60-80 kDa Rabbit IgG
DR3 (D4O3X) Rabbit mAb 20772 20 µl 55-60 kDa Rabbit IgG
DR4 (D9S1R) Rabbit mAb 42533 20 µl 35-55 kDa Rabbit IgG
DR5 (D4E9) XP® Rabbit mAb 8074 20 µl 40, 48 kDa Rabbit IgG
DR6 (E8D2I) Rabbit mAb 93026 20 µl 80, 120 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 Death Receptor Antibody Sampler Kit II provides an economical means to investigate members of the death receptor family. The 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

The tumor necrosis factor receptor family, which includes TNF-RI, TNF-R2, Fas, DR3, DR4, DR5, and DR6, plays an important role in the regulation of apoptosis in various physiological systems (1,2). The receptors are activated by a family of cytokines that include TNF, FasL, TWEAK, and TRAIL. They are characterized by a highly conserved extracellular region containing cysteine-rich repeats and a conserved intracellular region of about 80 amino acids termed the death domain (DD). The DD is important for transducing the death signal by recruiting other DD containing adaptor proteins (FADD, TRADD, RIP) to the death-inducing signaling complex (DISC) resulting in activation of caspases. The two receptors for TNF-α, TNF-R1 (55 kDa) and TNF-R2 (75 kDa) can mediate distinct cellular responses (3,4). In most cases cytotoxicity elicited by TNF has been reported to act through TNF-R1 (5,6). DR3/WSL-1/Apo-3/TRAMP/LARD is a TNFR family member containing the characteristic extracellular cysteine-repeats, transmembrane region, and an intracellular DD (7-11). DR3 is activated by its ligand Apo-3L/TWEAK to induce apoptosis and activation of NF-κB (12,13). Like TNF-R1, DR3 binds to the DD adaptor protein TRADD, which can then associate with other DD proteins like FADD and RIP as well as members of the TRAF family (7,8). Tissue expression of DR3 is very restricted, primarily seen on the surface of activated thymocytes and lymphocytes and plays an important role in thymocyte negative selection (7,8,14). Studies have also indicated an association with DR3 and rheumatoid arthritis (15,16). DR4 (TRAIL-RI, TNFRSF10A) and DR5 (TRAIL-R2, TNFRSF10B) are receptors for the cytokine TRAIL. Both receptors contain death domains that recruit DISC complexes triggering caspase activation and apoptosis (17-20). DR6, also known as TNFRSF21, is a TNFR family member able to induce apoptosis as well as activation of NF-κB and JNK (21). DR6 appears to play a critical role in the activation and differentiation of T and B lymphocytes (22,23). In the nervous system, β-amyloid precursor protein (APP) activates DR6 to trigger neuronal degeneration (24).

  1. Nagata, S. (1997) Cell 88, 355-65.
  2. Thorburn, A. (2004) Cell Signal 16, 139-44.
  3. Tartaglia, L.A. et al. (1991) Proc Natl Acad Sci U S A 88, 9292-6.
  4. Peschon, J.J. et al. (1998) J Immunol 160, 943-52.
  5. Tartaglia, L.A. et al. (1993) Cell 73, 213-6.
  6. Rothe, J. et al. (1993) Nature 364, 798-802.
  7. Chinnaiyan, A.M. et al. (1996) Science 274, 990-2.
  8. Kitson, J. et al. (1996) Nature 384, 372-5.
  9. Marsters, S.A. et al. (1996) Curr Biol 6, 1669-76.
  10. Bodmer, J.L. et al. (1997) Immunity 6, 79-88.
  11. Screaton, G.R. et al. (1997) Proc Natl Acad Sci U S A 94, 4615-9.
  12. Marsters, S.A. et al. (1998) Curr Biol 8, 525-8.
  13. Kaptein, A. et al. (2000) FEBS Lett 485, 135-41.
  14. Wang, E.C. et al. (2001) Mol Cell Biol 21, 3451-61.
  15. Osawa, K. et al. (2004) Genes Immun 5, 439-43.
  16. Borysenko, C.W. et al. (2005) Biochem Biophys Res Commun 328, 794-9.
  17. Pan, G. et al. (1997) Science 276, 111-3.
  18. Walczak, H. et al. (1997) EMBO J 16, 5386-97.
  19. Chaudhary, P.M. et al. (1997) Immunity 7, 821-30.
  20. Schneider, P. et al. (1997) Immunity 7, 831-6.
  21. Pan, G. et al. (1998) FEBS Lett 431, 351-6.
  22. Zhao, H. et al. (2001) J Exp Med 194, 1441-8.
  23. Schmidt, C.S. et al. (2003) J Exp Med 197, 51-62.
  24. Nikolaev, A. et al. (2009) Nature 457, 981-9.

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 4
    #29603

    Death Receptor Antibody Sampler Kit II

    Death Receptor Antibody Sampler Kit II: Image 1 Expand Image
    使用 TNF-R1 (C25C1) Rabbit mAb #3736 对 LN18 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。
    Death Receptor Antibody Sampler Kit II: Image 2 Expand Image
    使用 DR3 (D4O3X) Rabbit mAb(上)或 β-Actin (D6A8) Rabbit mAb #8457(下),对不同细胞系进行蛋白质印迹分析。KARPAS 细胞系来源:剑桥大学的 Abraham Kapas 博士。
    Death Receptor Antibody Sampler Kit II: Image 3 Expand Image
    使用 TNF-R1 (C25C1) Rabbit mAb 对 A549、LN-18 和 Hela 细胞提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 4 Expand Image
    使用 Fas (C18C12) Rabbit mAb 对转染空载或经人 Fas 转染的 COS 细胞以及 ACHN 和 HT-1080 细胞系的提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 5 Expand Image
    使用 DR4 (D9S1R) Rabbit mAb (上)或 β-Actin (D6A8) Rabbit mAb #8457(下),对来自不同细胞系的提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 6 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。
    Death Receptor Antibody Sampler Kit II: Image 7 Expand Image
    使用 TNF-R2 (E8D7P) Rabbit mAb(右图)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)和共染色的 NCAM1 (CD56) (MY31) Mouse mAb (APC Conjugate) #36377 对活性 人外周血单核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。
    Death Receptor Antibody Sampler Kit II: Image 8 Expand Image
    使用 DR5 (D4E9) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 9 Expand Image
    使用 DR6 (E8D2I) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对不同细胞系的提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 10 Expand Image
    使用 DR3 (D4O3X) Rabbit mAb ,对转染空载 (-) 或转染表达 Myc/DDK 标签的人全长DR3 (hDR3-Myc/DDK;+)的表达载体的 293T 细胞提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 11 Expand Image
    使用 Fas (C18C12) Rabbit mAb 对石蜡包埋的人结肠癌组织进行免疫组织化学分析。
    Death Receptor Antibody Sampler Kit II: Image 12 Expand Image
    使用 DR4 (D9S1R) Rabbit mAb,对转染空载 (-) 或转染表达人全长 DR4 (hDR4;+)的表达载体的 293T 细胞提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 13 Expand Image
    使用 TNF-R2 (E8D7P) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对不同人体细胞提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 14 Expand Image
    使用 DR5 (D4E9) XP® Rabbit mAb 对未经处理或经阿霉素 (500 nM) 处理指定时间的 A549 细胞提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 15 Expand Image
    使用 DR6 (E8D2I) Rabbit mAb 对转染空载 (-) 或转染表达全长人 DR6 (hDR6; +) 的表达载体的 293T 细胞的提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 16 Expand Image
    在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 Fas (C18C12) Rabbit mAb 对石蜡包埋的人结肠进行免疫组织化学分析。
    Death Receptor Antibody Sampler Kit II: Image 17 Expand Image
    对来自 PC-3 细胞提取物的DR4 进行免疫沉淀。泳道 1 表示 10 % input,泳道 2 是 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,并且泳道 3 是 DR4 (D9S1R) Rabbit mAb。使用 DR4 (D9S1R) Rabbit mAb 进行蛋白质印迹。使用构象特异性二抗来避免 IgG 交叉反应性。
    Death Receptor Antibody Sampler Kit II: Image 18 Expand Image
    对 SW620 细胞提取物 TNF-R2 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 TNF-R2 (E8D7P) Rabbit mAb。使用 TNF-R2 (E8D7P) Rabbit mAb 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
    Death Receptor Antibody Sampler Kit II: Image 19 Expand Image
    使用 DR5 (D4E9) XP® Rabbit mAb 对转染空载 (-) 或转染表达人 DR5 长同工型 (hDR5, +) 的表达载体的 293T 细胞提取物进行蛋白质印迹分析。
    Death Receptor Antibody Sampler Kit II: Image 20 Expand Image
    对 RT4 细胞提取物 DR6 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 DR6 (E8D2I) Rabbit mAb。使用 DR6 (E8D2I) Rabbit mAb 进行蛋白质印迹实验。Anti-Rabbit, HRP-linked Antibody #7074 用作二抗。
    Death Receptor Antibody Sampler Kit II: Image 21 Expand Image
    使用 Fas (C18C12) Rabbit mAb 对石蜡包埋的 Raji(阳性,左)或 K562 细胞(阴性,右)进行免疫组织化学分析。
    Death Receptor Antibody Sampler Kit II: Image 22 Expand Image
    使用 DR4 (D9S1R) Rabbit mAb(绿色),对 HCT 116(阳性,左)细胞和 FaDu(阴性,右)细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Death Receptor Antibody Sampler Kit II: Image 23 Expand Image
    使用 DR5 (D4E9) XP® Rabbit mAb(绿色)对 HT-1080 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Death Receptor Antibody Sampler Kit II: Image 24 Expand Image
    使用与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(红)相对比的 DR4 (D9S1R) Rabbit mAb(蓝),对固定和通透的 U266 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。