Revision 6
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, IF-F, FC-FP, FC-L

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

40-80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P01590

Entrez-Gene Id:

16184

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
Flow Cytometry (Live) 1:100 - 1:400

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 #95742.

Specificity / Sensitivity

IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb recognizes endogenous levels of total IL-2Rα/CD25 protein. This antibody non-specifically labels myenteric nerves in the gut.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of mouse IL-2Rα/CD25 protein.

Background

Interleukin-2 (IL-2) is a T cell stimulatory cytokine best known for inducing T cell proliferation and NK cell proliferation and activation (1,2). IL-2 also promotes peripheral development of regulatory T cells (Tregs) (3,4). Conversely, IL-2 is involved in the activation-induced cell death (AICD) that is observed post T cell expansion by increasing levels of Fas on CD4+ T cells (5). The effects of IL-2 are mediated through a trimeric receptor complex consisting of IL-2Rα, IL-2Rβ, and the common gamma chain, γc (1,2). IL-2Rα binds exclusively to IL-2 with low affinity and increases the binding affinity of the whole receptor complex including IL-2Rβ and γc subunits. IL-15 also binds to IL-2Rβ (1,2). γc is used by other cytokines, including IL-4, IL-7, IL-9, IL-15, and IL-21 (1,2). Binding of IL-2 initiates signaling cascades involving Jak1, Jak3, Stat5, and the PI3K/Akt pathways (1,2).

  1. Ma, A. et al. (2006) Annu Rev Immunol 24, 657-79.
  2. Gaffen, S.L. and Liu, K.D. (2004) Cytokine 28, 109-23.
  3. Fehérvari, Z. et al. (2006) Trends Immunol 27, 109-11.
  4. Antony, P.A. et al. (2006) J Immunol 176, 5255-66.
  5. Jaleco, S. et al. (2003) J Immunol 171, 61-8.

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 nonfat dry milk, 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) IF-F: Immunofluorescence (Frozen) FC-FP: Flow Cytometry (Fixed/Permeabilized) FC-L: Flow Cytometry (Live)

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

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 6
#39475

IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb

Western Blotting Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对不同细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
对 CTLL-2 细胞提取物中的 IL-2Rα/CD25 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb。使用 IL-2Rα/CD25 (E9W2J) 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: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb 对石蜡包埋的小鼠胸腺进行免疫组织化学分析。
Immunohistochemistry Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb 对石蜡包埋的 4T1 同源乳腺肿瘤进行免疫组织化学分析。
Immunohistochemistry Image 2: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb 对石蜡包埋的 A20 同源肿瘤进行免疫组织化学分析。
Immunohistochemistry Image 3: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb 对石蜡包埋的 CTLL-2 细胞沉淀物(左图,阳性)或 NIH/3T3 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunohistochemistry Image 4: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(左图)或 IL-2Rα/CD25 (E4B2Z) XP® Rabbit mAb #36128(右图)对石蜡包埋的小鼠结肠进行免疫组织化学分析。这两种抗体可检测小鼠 IL-2Rα/CD25 上的独立独特表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 5: IL-2Rα/CD25 (E9W2J) 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 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(左图)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。
Immunofluorescence Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(绿色)对小鼠脾细胞进行共聚焦免疫荧光分析。使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900封闭游离的二抗结合位点后,用 CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127(伪彩蓝色) 和 FoxP3 (D6O8R) Rabbit mAb (Alexa Fluor® 647 Conjugate)(红色)标记组织。
Immunofluorescence Image 2: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(绿色)、CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127(伪彩红色)和 CD45R/B220 (RA3-6B2) Rat mAb (redFluor 710 Conjugate) #82984(伪彩青色)对小鼠脾脏进行共聚焦免疫荧光分析。切片用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。
Immunofluorescence Image 3: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(伪彩绿色)和 CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127(伪彩红色)对小鼠结肠进行共聚焦免疫荧光分析。切片用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。注:肌层神经丛中的信号被认为是非特异性的。
Flow Cytometry Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(右图) 或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)和共染料 CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127 对活化小鼠脾细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 作为二抗。
Flow Cytometry Image 2: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
使用 IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对活化 C2C12 细胞(蓝色,阴性)和 CTLL-2 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。