Revision 4

#82000Store 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):

78

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q04864

Entrez-Gene Id:

5966

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800

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.

For a carrier free (BSA and azide free) version of this product see product #57603.

Specificity / Sensitivity

c-Rel (E8Z5Y) XP® Rabbit mAb recognizes endogenous levels of total c-Rel protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human c-Rel protein.

Background

Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).

c-Rel contains an amino-terminal DNA-binding domain, referred to as the REL homology domain (REH), and carboxy-terminal transactivation domains. The c-Rel protein is typically inhibited in unstimulated cells by IκBα and IκBβ. c-Rel expression is highest in hematopoietic cells with extensive research studies demonstrating its role in immune cell function and pathogenesis of disease (12,13).

  1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
  2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
  3. Haskill, S. et al. (1991) Cell 65, 1281-9.
  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
  8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  9. Senftleben, U. et al. (2001) Science 293, 1495-9.
  10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
  11. Xiao, G. et al. (2001) Mol Cell 7, 401-9.
  12. Gilmore, T.D. and Gerondakis, S. (2011) Genes Cancer 2, 695-711.
  13. Fullard, N. et al. (2012) Int J Biochem Cell Biol 44, 851-60.

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.
XP is a registered trademark of Cell Signaling Technology, Inc.
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
#82000

c-Rel (E8Z5Y) XP® Rabbit mAb

Western Blotting Image 1: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对来自各种细胞系的提取物进行蛋白质印迹分析。c-Rel 蛋白在 HL-60、MDA-MB-453 和 Caki-1 细胞中表达较少与预测的表达模式一致。
Immunoprecipitation Image 1: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
对 RT-4 细胞提取物的 c-Rel 蛋白进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 c-Rel (E8Z5Y) XP® Rabbit mAb。使用 c-Rel (E8Z5Y) XP® 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。
Immunohistochemistry Image 1: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人 T 细胞淋巴瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 2: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人宫颈鳞状细胞癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人尿路上皮癌组织进行免疫组织化学分析。
Immunohistochemistry Image 4: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人卵巢浆液性腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 5: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人导管乳腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 6: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人尿路上皮癌组织进行免疫组织化学分析。
Immunohistochemistry Image 7: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的人正常肾脏组织进行免疫组织化学分析。
Immunohistochemistry Image 8: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb 对石蜡包埋的 Ri-1 细胞沉淀物(左图,高表达)或 MDA-MB-453 细胞沉淀物(右图,低表达)进行免疫组织化学分析。
Immunohistochemistry Image 9: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)比较,使用 c-Rel (E8Z5Y) XP® Rabbit mAb(左图)对石蜡包埋的人正常淋巴结组织进行免疫组织化学分析。
Immunofluorescence Image 1: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色),对未经处理(左图)或经 hTNF-α(20 ng/mL,30 分钟;中图)处理的 RT-4 细胞(高表达)和 Caki-1 细胞(右图,低表达)进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: c-Rel (E8Z5Y) XP® Rabbit mAb Expand Image
使用 c-Rel (E8Z5Y) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 HL-60 细胞(蓝色,低表达)和 RT-4 细胞(绿色,高表达)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。