Revision 3
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.
REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Mouse IgG1 kappa

UniProt ID:

#P10747

Entrez-Gene Id:

940

Product Information

Formulation

Supplied in 10 mM NaH2PO4, 150 mM NaCl, pH7.2, Low Endotoxin, Azide-free.

Storage

Stable for 12 months when stored at 4°C. Do not aliquot the antibody.

Specificity / Sensitivity

Human CD28 Activating (CD28.2) Mouse mAb (Low Endotoxin, Azide-free) recognizes endogenous levels of CD28 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced using cells transfected with human CD28 and purified via affinity chromatography.

Product Description

Endotoxin Less than or equal to 0.01 EU/μg, as determined by the LaL assay.

Background

CD28 is a transmembrane glycoprotein expressed by T cells as well as some other hematopoietic cells (1, 2). T cell activation requires T cell receptor (TCR) recognition of antigen presented in the context of MHC molecules. CD28 acts as a T cell costimulatory receptor, and interaction of CD28 with its ligands CD80 or CD86 provides the second signal required for naïve T cell activation (3-5). Activation of naïve T cells in the absence of CD28 stimulation can result in a state of T cell anergy, or unresponsiveness (3). CD28 signals through cytoplasmic phospho-tyrosine motifs that bind several SH2 or SH3 domain-containing proteins involved in T cell activation (2). Recently, CD28 was demonstrated to be a preferred target of PD-1-mediated dephosphorylation. Consistently, CD28 expression was required for T cell proliferation following PD-1 blockade and CD28 stimulation was required for effective anti-PD-1 cancer immunotherapy in mice (6, 7). Several CD28 isoforms are produced by alternative splicing (8).

  1. Aruffo, A. and Seed, B. (1987) Proc Natl Acad Sci U S A 84, 8573-7.
  2. Esensten, J.H. et al. (2016) Immunity 44, 973-88.
  3. Harding, F.A. et al. (1992) Nature 356, 607-9.
  4. Azuma, M. et al. (1993) Nature 366, 76-9.
  5. Linsley, P.S. et al. (1990) Proc Natl Acad Sci U S A 87, 5031-5.
  6. Hui, E. et al. (2017) Science 355, 1428-1433.
  7. Kamphorst, A.O. et al. (2017) Science 355, 1423-1427.
  8. Magistrelli, G. et al. (1999) Biochem Biophys Res Commun 259, 34-7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

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 3
#91920

Human CD28 Activating (CD28.2) Mouse mAb (Low Endotoxin, Azide-free)

Human CD28 Activating (CD28.2) Mouse mAb (Low Endotoxin, Azide-free): Image 1 Expand Image
使用 Phospho-SLP-76 (Ser376) (E3G9U) XP® Rabbit mAb #76384(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(蓝色)或经过交联的 Human CD3ε Activating (OKT3) Mouse mAb (Low Endotoxin, Azide-free) #92511和 Human CD28 Activating (CD28.2) Mouse mAb (Low Endotoxin, Azide-free)(各为 10 μg/ml,15 分钟;绿色)处理的 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Human CD28 Activating (CD28.2) Mouse mAb (Low Endotoxin, Azide-free): Image 2 Expand Image
使用 Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb #92711(上图)、SLP-76 (E4N7E) Rabbit mAb #25361(中图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对未经处理 (1) 或经过交联的 Human CD3ε Activating (OKT3) Mouse mAb (Low Endotoxin, Azide-free) #92511 和 Human CD28 Activating (CD28.2) Mouse mAb (Low Endotoxin, Azide-free)(10 μg/ml,30 分钟)(2) 处理的 Jurkat 细胞提取物进行蛋白质印迹分析。