Revision 5

#67223Store 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-F

REACTIVITY:

M R

SENSITIVITY:

Endogenous

MW (kDa):

22-35

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9WVS0

Entrez-Gene Id:

54167

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 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 #18670.

Specificity / Sensitivity

ICOS (E5F7E) Rabbit mAb recognizes endogenous levels of total mouse ICOS protein.

Species Reactivity:

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val198 of mouse ICOS protein.

Background

ICOS (Inducible Co-Stimulator, CD278) is a member of the CD28 family that regulates T cell activity and immune responses (1). The ICOS protein contains an extracellular IgV-like domain, a transmembrane domain, and an intracellular domain with a YMFM motif (1-2). ICOS is primarily expressed on activated CD4+ and CD8+ T cells (1). Upon binding to its ligand, ICOS potentiates the T cell response to antigen through activation of the PI3K signaling pathway (3). In addition to enhancing T cell activation and proliferation, ICOS plays an important role in the regulation of T follicular helper cells (4). Research studies suggest that ICOS is a potential therapeutic target, and could serve as a prognostic biomarker for neoplastic therapy involving CTLA-4 blockade (5-7).

  1. Hutloff, A. et al. (1999) Nature 397, 263-6.
  2. Coyle, A.J. et al. (2000) Immunity 13, 95-105.
  3. Fos, C. et al. (2008) J Immunol 181, 1969-77.
  4. Hutloff, A. (2015) Oncotarget 6, 21785-6.
  5. Kyi, C. and Postow, M.A. (2014) FEBS Lett 588, 368-76.
  6. Fan, X. et al. (2014) J Exp Med 211, 715-25.
  7. Ng Tang, D. et al. (2013) Cancer Immunol Res 1, 229-34.

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-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen)

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 5
#67223

ICOS (E5F7E) Rabbit mAb

Western Blotting Image 1: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对不同组织和细胞的提取物进行蛋白印迹分析。CD4+ T 细胞用小鼠脾或人血细胞纯化而来,并在添加 Mouse Interleukin-2 (mIL-2) (20 ng/ml) 或 Human Interleukin-2 (hIL-2) #8907 (20 ng/ml) 的情况下,用涂有 CD3 和 CD28 抗体的珠子刺激 7 天。
Immunoprecipitation Image 1: ICOS (E5F7E) Rabbit mAb Expand Image
对小鼠 CD4+ T 细胞提取物 ICOS 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 ICOS (E5F7E) Rabbit mAb。使用 ICOS (E5F7E) Rabbit mAb 进行蛋白印迹分析。CD4+ T 细胞纯化自小鼠脾脏,并在存在 Mouse Interleukin-2 (mIL-2) (20 ng/ml) 的情况下,用涂有 CD3 和 CD28 抗体的珠子刺激 7 天。
Immunohistochemistry Image 1: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb 对石蜡包埋的正常小鼠胸腺进行免疫组织化学分析。
Immunohistochemistry Image 2: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb 对石蜡包埋的正常小鼠脾脏进行免疫组织化学分析。
Immunohistochemistry Image 3: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb 对石蜡包埋的正常小鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 4: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb 对石蜡包埋的 4T1 同源乳腺肿瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 5: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) 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 6: ICOS (E5F7E) 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 ICOS (E5F7E) Rabbit mAb(左图)对石蜡包埋的正常小鼠脾脏进行免疫组织化学分析。
Immunohistochemistry Image 7: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb 对石蜡包埋的纯化 CD4+ 细胞沉淀物(左图,阳性)或 C2C12 细胞沉淀物(右图,阴性)进行免疫组织化学分析。CD4+ T 细胞纯化自小鼠脾脏,并在存在 Mouse Interleukin-2 (mIL-2) (20 ng/ml) 的情况下,用涂有 CD3 和 CD28 抗体的珠子刺激 12 天。
Immunofluorescence Image 1: ICOS (E5F7E) Rabbit mAb Expand Image
使用 ICOS (E5F7E) Rabbit mAb(伪彩绿色)和 CD4 (RM4-5) Rat mAb #96127 (FITC Conjugate)(伪彩红色)对小鼠脾脏进行共聚焦免疫荧光分析。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。