Revision 1
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:

IF-IC, FC-FP, FC-L

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9Y5U5

Entrez-Gene Id:

8784

Product Information

Product Usage Information

Application Dilution
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:50
Flow Cytometry (Live) 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

GITR (D5V7P) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total GITR protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human GITR protein.

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric and immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated GITR (D5V7P) Rabbit mAb #10419.

Background

TNFRSF18, also known as glucocorticoid-induced tumor necrosis factor-receptor (TNFR)-related protein (GITR) and activation-inducible TNFR family receptor, encodes a type 1 membrane protein of the TNF-receptor superfamily (1). Three alternatively spliced transcript variants encoding distinct isoforms have been reported (2). GITR is an immune cell co-stimulatory receptor expressed constitutively at high levels on CD4+ CD25+ T regulatory cells (Tregs), at low levels on naïve and memory T cells, and is induced upon T cell activation (3-5). Studies show GITR can also be induced on NK cells, macrophages, and DCs (3,4,6). Although GITR does not have intrinsic enzymatic activity, TNFSF18 (also known as GITRL) expressed on antigen presenting cells binds to GITR, resulting in recruitment of TNFR-associated factor family members and activation of the NF-κB pathway in T cells (7). GITR ligation has been shown to play a role in CD8+ T cell activation, cytotoxicity, and memory T cell survival (8-10). In the thymus, GITR is thought to play a key role in dominant immunological self-tolerance through thymic Treg differentiation and expansion (11). Of note, GITR ligation inhibits Treg suppressive function (12-13) and promotes effector T cell resistance to Treg suppression (14-15). Due to the combined effects on both Treg suppression and effector cell activation, GITR represents a unique opportunity for immunotherapeutic intervention in cancer (16).

  1. Nocentini, G. et al. (1997) Proc Natl Acad Sci U S A 94, 6216-21.
  2. Nocentini, G. et al. (2000) Cell Death Differ 7, 408-10.
  3. Shimizu, J. et al. (2002) Nat Immunol 3, 135-42.
  4. Nocentini, G. and Riccardi, C. (2009) Adv Exp Med Biol 647, 156-73.
  5. McHugh, R.S. et al. (2002) Immunity 16, 311-23.
  6. Hanabuchi, S. et al. (2006) Blood 107, 3617-23.
  7. Snell, L.M. et al. (2011) Immunol Rev 244, 197-217.
  8. Ronchetti, S. et al. (2007) J Immunol 179, 5916-26.
  9. Kim, I.K. et al. (2015) Nat Med 21, 1010-7.
  10. Snell, L.M. et al. (2012) J Immunol 188, 5915-23.
  11. Petrillo, M.G. et al. (2015) Autoimmun Rev 14, 117-26.
  12. Kanamaru, F. et al. (2004) J Immunol 172, 7306-14.
  13. Valzasina, B. et al. (2005) Blood 105, 2845-51.
  14. Stephens, G.L. et al. (2004) J Immunol 173, 5008-20.
  15. Nishikawa, H. et al. (2008) Cancer Res 68, 5948-54.
  16. Knee, D.A. et al. (2016) Eur J Cancer 67, 1-10.

Species Reactivity

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

Applications Key

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.
XP is a registered trademark of Cell Signaling Technology, Inc.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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Revision 1
#65705

GITR (D5V7P) Rabbit mAb (Alexa Fluor® 488 Conjugate)

Immunofluorescence Image 1: GITR (D5V7P) Rabbit mAb (Alexa Fluor® 488 Conjugate) Expand Image
使用 GITR (D5V7P) Rabbit mAb (Alexa Fluor® 488 Conjugate)(绿色)对 HuT 102 细胞(左图,阳性)和 Jurkat 细胞(右图,阴性)进行共聚焦免疫荧光分析。肌动蛋白丝已带有 Alexa Fluor® 647 Phalloidin #8940(红色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Flow Cytometry Image 1: GITR (D5V7P) Rabbit mAb (Alexa Fluor® 488 Conjugate) Expand Image
使用 GITR (D5V7P) Rabbit mAb (Alexa Fluor® 488 Conjugate) (顶行)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975(底行)和共染料 CD3 (UCHT1) Mouse mAb (APC Conjugate) #19881 对未经处理(左栏)或已经过抗 CD3(10 μg/ml,72 小时)和抗 CD28(5 μg/ml,72 小时)处理的人外周血单核细胞进行流式细胞分析。