Revision 1

#66102Store 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, IF-F, IF-IC, FC-FP, FC-L

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

100-200

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q3U0X8

Entrez-Gene Id:

84544

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Frozen) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200
Flow Cytometry (Live) 1:50 - 1:200

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

CD96 (E1Y2Z) Rabbit mAb recognizes endogenous levels of total CD96 protein. This antibody does not cross-react with human CD96 protein.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the extracellular domain of mouse CD96 protein.

Background

Cluster of Differentiation 96 (CD96), also known as T cell-activated increased late expression protein (TACTILE), is a heavily glycosylated type I transmembrane protein belonging to the immunoglobulin superfamily (1-3). CD96 is expressed on peripheral T cells and natural killer (NK) cells, and is strongly upregulated on these cell types after activation (1,4). It is also expressed on transformed T cells, and at very low levels on activated B cells (1,5,6). CD96 interacts with the receptor ligand PVR (CD155), and is involved in adhesion of activated T and NK cells to target cells during the late phase of the immune response (2,4). CD96 competes with DNAM-1 (CD226) for binding to PVR (CD155) in order to negatively regulate T and NK cell activity, and thus blockade of CD96 in order to induce anti-tumor immune responses is investigated for potential immunotherapeutic intervention in cancer (7-9). Alternative splicing occurs at this locus, and two transcript variants encoding distinct isoforms with variable functions have been identified (1,10).

  1. Wang, P.L. et al. (1992) J Immunol 148, 2600-8.
  2. Seth, S. et al. (2007) Biochem Biophys Res Commun 364, 959-65.
  3. Georgiev, H. et al. (2018) Front Immunol 9, 1072.
  4. Fuchs, A. et al. (2004) J Immunol 172, 3994-8.
  5. Burger, R. et al. (1999) Leuk Res 23, 19-27.
  6. Gramatzki, M. et al. (1998) Exp Hematol 26, 1209-14.
  7. Chan, C.J. et al. (2014) Nat Immunol 15, 431-8.
  8. Blake, S.J. et al. (2016) Cancer Discov 6, 446-59.
  9. Brooks, J. et al. (2018) Cancer Res 78, 475-88.
  10. Meyer, D. et al. (2009) J Biol Chem 284, 2235-44.

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 IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) 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.
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 1
#66102

CD96 (E1Y2Z) Rabbit mAb

Western Blotting Image 1: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对来自 EL4、CTLL-2 和 C2C12 细胞的提取物进行蛋白质印迹法分析。CD96 蛋白在 C2C12 细胞中的阴性表达与预测的表达模式一致。
Western Blotting Image 2: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(上图)、Myc-Tag (71D10) Rabbit mAb #2278(中图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载 (-) 或转染带有 Myc/DDK 标签的全长小鼠 CD96 (mCD96-Myc/DDK; +) 和带 Myc/DDK 标签的全长人 CD96 (hCD96-Myc/DDK; +) 表达载体的 293T 细胞提取物进行蛋白质印迹法分析。
Immunofluorescence Image 1: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(绿色)、CD45R/B220 (RA3-6B2) Rat mAb (FITC Conjugate) #34399(伪彩品红色)和 CD3 (17A2) Rat mAb (APC Conjugate) #24265(红色)对固定冷冻的小鼠脾脏进行共聚焦免疫荧光分析。
Immunofluorescence Image 2: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(绿色)、CD45R/B220 (RA3-6B2) Rat mAb (FITC Conjugate) #34399(伪彩品红色)和 CD3 (17A2) Rat mAb (APC Conjugate) #24265(红色)对固定冷冻的小鼠淋巴结进行共聚焦免疫荧光分析。
Immunofluorescence Image 1: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(绿色)和 DAPI #4083(蓝色)对 EL4 细胞(左图,阳性)或 C2C12 细胞(右图,阴性)进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(右图)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)和共染色的 CD3 (17A2) Rat mAb (violetFluor 450 Conjugate) #38527 对活性小鼠脾脏进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Flow Cytometry Image 2: CD96 (E1Y2Z) Rabbit mAb Expand Image
使用 CD96 (E1Y2Z) Rabbit mAb(右图)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)和共染色的 CD3 (17A2) Rat mAb (violetFluor 450 Conjugate) #38527 对固定和通透的小鼠脾脏进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。