Revision 4

#11904Store 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, W-S, IP, IF-IC

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

50-55

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q14653

Entrez-Gene Id:

3661

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 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.

Specificity / Sensitivity

IRF-3 (D6I4C) XP® Rabbit mAb recognizes endogenous levels of total IRF-3 protein.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant human IRF-3 protein.

Background

Interferon regulatory factors (IRFs) comprise a family of transcription factors that function within the Jak/Stat pathway to regulate interferon (IFN) and IFN-inducible gene expression in response to viral infection (1). IRFs play an important role in pathogen defense, autoimmunity, lymphocyte development, cell growth, and susceptibility to transformation. The IRF family includes nine members: IRF-1, IRF-2, IRF-9/ISGF3γ, IRF-3, IRF-4 (Pip/LSIRF/ICSAT), IRF-5, IRF-6, IRF-7, and IRF-8/ICSBP. All IRF proteins share homology in their amino-terminal DNA-binding domains. IRF family members regulate transcription through interactions with proteins that share similar DNA-binding motifs, such as IFN-stimulated response elements (ISRE), IFN consensus sequences (ICS), and IFN regulatory elements (IRF-E) (2).

IRF-3 can inhibit cell growth and plays a critical role in controlling the expression of genes in the innate immune response (1-4). In unstimulated cells, IRF-3 is present in the cytoplasm. Viral infection results in phosphorylation of IRF-3 and leads to its translocation to the nucleus where it activates promoters containing IRF-3-binding sites. Phosphorylation of IRF-3 occurs at a cluster of C-terminal Ser and Thr residues (between 385 and 405), leading to its association with the p300/CBP coactivator protein that promotes DNA binding and transcriptional activity (5). During infection, IRF-3 is likely activated through a pathway that includes activation of Toll-like receptors and a kinase complex that includes IKKε and TBK1 (6,7). IRF-3 is phosphorylated at Ser396 following viral infection, expression of viral nucleocapsid, and double-stranded RNA treatment. These events likely play a role in activation of IRF-3 (8).

  1. Taniguchi, T. et al. (2001) Annu Rev Immunol 19, 623-55.
  2. Honda, K. and Taniguchi, T. (2006) Nat Rev Immunol 6, 644-58.
  3. Hiscott, J. et al. (1999) J Interferon Cytokine Res 19, 1-13.
  4. Kim, T.Y. et al. (2003) J Biol Chem 278, 15272-8.
  5. Yoneyama, M. et al. (2002) J Interferon Cytokine Res 22, 73-6.
  6. Fitzgerald, K.A. et al. (2003) Nat Immunol 4, 491-6.
  7. Kopp, E. and Medzhitov, R. (2003) Curr Opin Immunol 15, 396-401.
  8. Servant, M.J. et al. (2003) J Biol Chem 278, 9441-7.

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 W-S: Simple Western™ IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)

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

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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 4
#11904

IRF-3 (D6I4C) XP® Rabbit mAb

Western Blotting Image 1: IRF-3 (D6I4C) XP® Rabbit mAb Expand Image
使用 IRF-3 (D6I4C) XP® Rabbit mAb, #11904(上图)或 β-actin (13E5) Rabbit mAb, #4970(下图)对对照型 HeLa 细胞(泳道 1) 或 IRF-3 敲除型 HeLa 细胞(泳道 2)的提取物进行蛋白质印迹分析。IRF-3 敲除型 HeLa 细胞中没有信号,这证实了该抗体对 IRF-3 的特异性。
Western Blotting Image 2: IRF-3 (D6I4C) XP® Rabbit mAb Expand Image
使用 IRF-3 (D6I4C) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
Western Blotting Image 1: IRF-3 (D6I4C) XP® Rabbit mAb Expand Image
使用 IRF-3 (D6I4C) XP ® Rabbit mAb #11904 对 HeLa 细胞的裂解物 (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 1:50 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。
Immunoprecipitation Image 1: IRF-3 (D6I4C) XP® Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道 2)或 IRF-3 (D6I4C) XP® Rabbit mAb(泳道 3),对 THP-1 细胞提取物的 IRF-3进行免疫沉淀。泳道 1 是 10% 输入对照。使用 IRF-3 (D6I4C) XP® Rabbit mAb 进行蛋白质印迹分析。使用 Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 和 Anti-mouse IgG, HRP-linked Antibody #7076 作为二抗。
Immunofluorescence Image 1: IRF-3 (D6I4C) XP® Rabbit mAb Expand Image
使用 IRF-3 (D6I4C) XP® Rabbit mAb(绿色)对未经(左)或经 Poly (I:C)(2.5 μg/ml,6 小时;右)转染的 HT-29 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。