Revision 4

#62834Store 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-Bond, IHC-P, FC-FP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

51

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q15306

Entrez-Gene Id:

3662

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 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 #56471.

Specificity / Sensitivity

IRF-4 (E8H3S) XP® Rabbit mAb recognizes endogenous levels of total IRF-4 protein.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to full-length human IRF-4 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-4 was independently cloned by three groups and demonstrated to have roles in different contexts of lymphoid regulation (3-5). First, IRF-4 (Pip) was found to associate with PU.1, a hematopoietic specific member of the ETS family, and to regulate the expression of B-cell specific genes (3). Second, it was characterized as a lymphoid-specific member of the IRF family (LSIRF) able to bind to ISRE (4). Third, it was identified in activated T cells as a factor that binds to the promoter of the interleukin-5 gene (ICSAT), and it was shown to repress gene activation induced by IFN (5). IRF-4 is expressed in all stages of B cell development and in mature T cells, and is inducible in primary lymphocytes by antigen mimetic stimuli such as concavalin A, CD3 crosslinking, anti-IgM and PMA treatment (4,5). Mice deficient in IRF-4 show normal distribution of B and T lymphocytes at 4 to 5 weeks, but later develop progressive generalized lymphadenopathy, suggesting a role for IRF-4 in the function and homeostasis of mature B- and T-lymphocytes (6).

  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. Eisenbeis, C.F. et al. (1995) Genes Dev 9, 1377-87.
  4. Matsuyama, T. et al. (1995) Nucleic Acids Res 23, 2127-36.
  5. Yamagata, T. et al. (1996) Mol Cell Biol 16, 1283-94.
  6. Mittrücker, H.W. et al. (1997) Science 275, 540-3.

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-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) 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.
KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
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
#62834

IRF-4 (E8H3S) XP® Rabbit mAb

Western Blotting Image 1: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb(上图)

与 β-Actin (D6A8) Rabbit mAb #8457(下图),对不同细胞系提取物进行蛋白质印迹分析。正如预计的一样,IRF-4 蛋白在 Jurkat 细胞中不表达。KARPAS-299 细胞系来源:剑桥大学 Abraham Karpas 博士。

Western Blotting Image 2: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb(上图)

与 β-Actin (D6A8) Rabbit mAb #8457(下图),对不同细胞系提取物进行蛋白质印迹分析。正如预计的一样,IRF-4 蛋白在 NIH/3T3 细胞中不表达。

Western Blotting Image 3: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对人原代 CD4 + T 细胞的提取物进行蛋白印迹分析。从人 PBMC 中纯化 CD4+ T 细胞,然后不处理(-)或使用涂有 CD3 和 CD28 抗体(+)的珠子处理 3 天。正如预计的一样,T 细胞激活可诱导 IRF-4 蛋白的上调。
Western Blotting Image 4: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb(上图)、DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368(中图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载 (-) 或转染表达带有 Myc/DDK 标记的全长人 IRF-4 蛋白 (hHLA-G-MycDDK; +) 的表达载体的 293T 细胞的提取物进行蛋白印迹分析。
Immunoprecipitation Image 1: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
对 Ramos 细胞提取物的 IRF-4 进行免疫沉淀。泳道 1 为 10 % input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 IRF-4 (E8H3S) XP® Rabbit mAb。使用 IRF-4 (E8H3S) XP® Rabbit mAb 进行蛋白印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP conjugate) #5127 用作二抗,

以避免与 IgG 发生交叉反应。

Immunohistochemistry Image 1: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的人鳞状细胞肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的人 T 细胞淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 1: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的正常人结肠进行免疫组织化学分析。
Immunohistochemistry Image 3: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 IRF-4 (E8H3S) XP® Rabbit mAb(左图)对石蜡包埋的人肺腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的小鼠肺 4T1 转移性肿瘤进行免疫组织化学分析。
Immunohistochemistry Image 5: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的小鼠小肠进行免疫组织化学分析。
Immunohistochemistry Image 6: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的 Ramos 细胞沉淀物(左图,阳性)或 Jurkat 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunohistochemistry Image 7: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb 对石蜡包埋的人扁桃体进行免疫组织化学分析。
Flow Cytometry Image 1: IRF-4 (E8H3S) XP® Rabbit mAb Expand Image
使用 IRF-4 (E8H3S) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 Jurkat 细胞(蓝色,阴性)和 Ramos 细胞(绿色,阳性)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。