Revision 3

#76684Store 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, ChIP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

48

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q00978

Entrez-Gene Id:

10379

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:3200
Chromatin IP 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

IRF-9 (D2T8M) Rabbit mAb recognizes endogenous levels of total IRF-9 protein. This antibody also cross-reacts with an unidentified protein of 95 kDa. Non-specific nucleolar background has been observed by immunofluorescence in some cultured cell lines.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to human IRF-9 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).

In response to type I interferons (e.g., IFN- α, IFN-β), interferon regulatory factor 9 (IRF-9, ISGF3γ) assembles into a protein complex with Stat1 and Stat2 and translocates to the nucleus where it induces transcription of interferon response genes by binding to ISREs (3,4). Expression of IRF-9 is also induced by type I interferons (5).

  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. Fu, X.Y. et al. (1990) Proc Natl Acad Sci U S A 87, 8555-9.
  4. Qureshi, S.A. et al. (1995) Proc Natl Acad Sci U S A 92, 3829-33.
  5. Levy, D.E. et al. (1990) EMBO J 9, 1105-11.

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) ChIP: Chromatin IP

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.
SimpleChIP is a registered 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 3
#76684

IRF-9 (D2T8M) Rabbit mAb

Western Blotting Image 1: IRF-9 (D2T8M) Rabbit mAb Expand Image
使用 IRF-9 (D2T8M) Rabbit mAb(上)或 β-Actin (D6A8) Rabbit mAb # 8457(下)对未处理(-) 或用 Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 小时; +) 处理的 HeLa 细胞进行蛋白质印迹分析。
Western Blotting Image 2: IRF-9 (D2T8M) Rabbit mAb Expand Image
使用 IRF-9 (D2T8M) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: IRF-9 (D2T8M) Rabbit mAb Expand Image
对 THP-1 细胞提取物中的 IRF-9 进行免疫沉淀。泳道 1 是 10% 的输入,泳道 2 则是 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,而泳道 3 是 IRF-9 (D2T8M) Rabbit mAb。使用 IRF-9 (D2T8M) Rabbit mAb 进行蛋白质印迹分析。
Immunofluorescence Image 1: IRF-9 (D2T8M) Rabbit mAb Expand Image
使用 IRF-9 (D2T8M) Rabbit mAb(绿色)对未处理(左) 或经 Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 小时; 右) 处理的 HeLa 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。
Chromatin Immunoprecipitation Image 1: IRF-9 (D2T8M) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 Human Interferon-α1 (hIFN-α1) # #8927(10 nM,30 分钟)处理的 U266 细胞的交联染色质和 IRF-9 (D2T8M) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Human MX1 Promoter Primers #57949 、human WARS intron 1 primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 方法对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。