Revision 1

#38916Store 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, IHC-P, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

27

Source/Isotype:

Rabbit IgG

UniProt ID:

#O60936

Entrez-Gene Id:

8996

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:400
Immunofluorescence (Immunocytochemistry) 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:100

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

ARC (D7Q3G) Rabbit mAb recognizes endogenous levels of total ARC protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro125 of human ARC protein, specific to a region encoded by isoform 2 of the NOL3 gene.

Background

Apoptosis repressor with caspase recruitment domain (ARC), also independently identified as muscle-enriched cytoplasmic protein (MYP), is a CARD domain protein that regulates apoptosis (1). The ARC protein CARD domain is highly homologous to those in other cell death regulators, including caspase-2, caspase-9, RAIDD, and Apaf-1 (2). The NOL3 gene encodes both the cytoplasmic ARC protein and a 30 kDa nucleolar protein (Nop30) that is involved in RNA splicing. ARC is encoded from isoform 2 of NOL3, while isoform 1 produced by alternative splicing encodes Nop30. Both ARC and Nop30 proteins share common amino-terminal sequences (3). Research studies show that ARC can bind to caspase-8 and caspase-2 and inhibit apoptosis through extrinsic pathways that involve the receptor proteins Fas, TNFR1, and DR3 (1). Additional research indicates that the ARC anti-apoptotic mechanism may include both intrinsic (mitochondrial) and extrinsic (death receptor) pathways (4). In addition to binding caspases, ARC can disrupt the interaction with the death domains of Fas and FADD, which inhibits death-inducing signaling complex (DISC) assembly. The CARD domain of ARC can inhibit intrinsic apoptosis through binding to the pro-apoptotic Bax protein (5). Phosphorylation of ARC at Thr149 by CK2 is required for targeting of ARC to the mitochondria (6). ARC is able to suppress necroptosis, a programmed pathway of necrosis triggered by blocking the recruitment of RIP1 to TNFR1 (7). Expression of ARC protein is predominantly seen in terminally differentiated cells under normal conditions and is markedly induced in a variety of cancers including pancreatic, colorectal, breast, lung, glioblastoma, liver, kidney, melanoma, and acute myeloid leukemia (1, 8-12).

  1. Koseki, T. et al. (1998) Proc Natl Acad Sci U S A 95, 5156-60.
  2. Hofmann, K. et al. (1997) Trends Biochem Sci 22, 155-6.
  3. Stoss, O. et al. (1999) J Biol Chem 274, 10951-62.
  4. Nam, Y.J. et al. (2004) Mol Cell 15, 901-12.
  5. Gustafsson, A.B. et al. (2004) J Biol Chem 279, 21233-8.
  6. Li, P.F. et al. (2002) Mol Cell 10, 247-58.
  7. Kung, G. et al. (2014) Cell Death Differ 21, 634-44.
  8. Mercier, I. et al. (2008) Cell Cycle 7, 1640-7.
  9. Wang, M. et al. (2005) FEBS Lett 579, 2411-5.
  10. Mercier, I. et al. (2005) Cell Death Differ 12, 682-6.
  11. Chen, L.H. et al. (2008) Cancer Res 68, 834-42.
  12. Carter, B.Z. et al. (2011) Blood 117, 780-7.

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 IHC-P: Immunohistochemistry (Paraffin) 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

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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

Revision 1
#38916

ARC (D7Q3G) Rabbit mAb

Western Blotting Image 1: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3G) 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: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3G) Rabbit mAb 对来自 293T 细胞的提取物进行蛋白质印迹分析。这些细胞是经转染试剂对照的 (-) 或经可表达

带 Myc/DDK 标签的人全长 Nop30 蛋白(hNop30-Myc/DDK;+)或带 Myc/DDK 标签的人全长 ARC 蛋白(hARC-Myc/DDK;+)的构建体转染的。

Immunoprecipitation Image 1: ARC (D7Q3G) Rabbit mAb Expand Image
对来自 MCF7 细胞提取物的 ARC 进行免疫沉淀。泳道 1 是 10% input,泳道 2 是 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,而泳道 3 是 ARC (D7Q3G) Rabbit mAb。使用 ARC (D7Q3G) Rabbit mAb 进行蛋白质印迹。构象特异性二抗用于避免与 IgG 的交叉反应性。
Immunohistochemistry Image 1: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3G) Rabbit mAb 对石蜡包埋的人淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 2: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3G) Rabbit mAb 对石蜡包埋的人卵巢癌进行免疫组织化学分析。
Immunohistochemistry Image 3: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3G) Rabbit mAb 对石蜡包埋的 MCF7(左)细胞沉淀物和 Daudi(右)细胞沉淀物进行免疫组织化学分析。
Immunohistochemistry Image 4: ARC (D7Q3G) Rabbit mAb Expand Image
在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 ARC (D7Q3G) Rabbit mAb 对石蜡包埋的人肺癌进行免疫组织化学分析。
Immunofluorescence Image 1: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3C) Rabbit mAb(绿色)对 MCF-7(阳性,左)细胞和 Daudi(阴性,右)细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。蓝色伪彩 = DRAQ5®#4084(DNA 荧光染料)。
Flow Cytometry Image 1: ARC (D7Q3G) Rabbit mAb Expand Image
使用 ARC (D7Q3G) Rabbit mAb 对 Daudi 细胞(蓝色)和 MCF7 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。