Revision 1

#42533Store at -20C

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.
Applications:

WB, IP, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

35-55

Source/Isotype:

Rabbit IgG

UniProt ID:

#O00220

Entrez-Gene Id:

8797

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50

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.

Specificity / Sensitivity

DR4 (D9S1R) Rabbit mAb recognizes endogenous levels of total DR4 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminal, cytoplasmic domain of human DR4 protein.

Background

The tumor necrosis factor receptor family, which includes TNF-RI, 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, and TNF-related apoptosis-inducing ligand (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.
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 (3-6). The ability of TRAIL to selectively kill malignant cells has led to clinical studies involving TRAIL and receptor agonists (7).

  1. Nagata, S. (1997) Cell 88, 355-65.
  2. Thorburn, A. (2004) Cell Signal 16, 139-44.
  3. Pan, G. et al. (1997) Science 276, 111-3.
  4. Walczak, H. et al. (1997) EMBO J 16, 5386-97.
  5. Chaudhary, P.M. et al. (1997) Immunity 7, 821-30.
  6. Schneider, P. et al. (1997) Immunity 7, 831-6.
  7. Yang, A. et al. (2010) Curr Opin Cell Biol 22, 837-44.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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
#42533

DR4 (D9S1R) Rabbit mAb

Western Blotting Image 1: DR4 (D9S1R) Rabbit mAb 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。
Western Blotting Image 2: DR4 (D9S1R) Rabbit mAb 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。
Immunoprecipitation Image 1: DR4 (D9S1R) Rabbit mAb 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。
Immunofluorescence Image 1: DR4 (D9S1R) Rabbit mAb 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)。
Flow Cytometry Image 1: DR4 (D9S1R) Rabbit mAb 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。