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In Vivo Ready™ Anti-Human CD3 (UCHT1) Antibody
- SPECIFICATION
- CITATIONS
- PROTOCOLS
- BACKGROUND
Application 
| WB, IHC-F, IF, FC, IP, FA |
|---|---|
| Isotype | Mouse IgG1, kappa |
| Concentration | 2 mg/mL |
| Reactivity | Human |
| Formulation | 10 mM NaH2PO4, 150 mM NaCl, pH7.2 |
| Host | Mouse |
| Gene ID | 915 |
|---|---|
| Gene Name | CD3D |
| Alternative Name(s) | Leu-4, T3 |
| Format | In Vivo Ready™ |
| Preparation | This monoclonal antibody preparation was purified from tissue culture supernatant via affinity chromatography. For In Vivo Ready™ (IVR) products, each preparation is also evaluated for endotoxin levels using the LAL assay. It is recommended to store the product undiluted at 4°C. Do not freeze. |
| Application Notes | This purified format is guaranteed to be >90% pure as determined by SDS-PAGE analysis. Citations are provided as a convenience to you - please consult Materials and Methods sections for additional details about the use of any product in these publications. |
| Endotoxin Level | Less than or equal to 0.01 EU/ug, as determined by the LaL assay |
| Storage Conditions | 2-8°C |
Harris SJ, Parry RV, Foster JG, Blunt MD, Wang A, Marelli-Berg F, Westwick J, and Ward SG. Apr. 2011. J. Immunol. 186: 4936-4945. (in vitro activation)
Beriou G, Bradshaw EM, Lozano E, Costantino CM, Hastings WD, Orban T, Elyaman W, Khoury SJ, Kuchroo VK, Baecher-Allan C, and Hafler DA. 2010. J. Immunol. 185: 46-54. (in vitro activation)
Soto PC, Stein LL, Hurtado-Ziola N, Hedrick SM, and Varki A. 2010. J. Immunol. 184: 4185-4195. (Flow cytometry – Chimpanzee)
Edelbauer M, Datta D, Vos IHC, Basu A, Stack MP, Reinders MEJ, Sho M, Calzadilla K, Ganze P, and Briscoe DM. 2010. Blood. 116:1980-1989. (Immunohistochemistry – acetone fixed, frozen sections; Immunofluoresence microscopy)
Varghese JC and Kane KP. 2008. J. Immunol. 181: 6002-6009. (in vitro activation)
Mack CL, Tucker RM, Sokol RJ, Darrer FM, Kotzin BL, Whitington PF and Miller SD. 2004. Pediatr. Res. 56(1):79-87. (Immunohistochemistry – frozen tissue)
Sakkas LI, Scanzello C, Johanson N, Burkholder J, Mitra A, Salgame P, Katsetos CD, and Platsoucas CD. 1998. Clin. Diagn. Lab. Immun. 5:430. (Immunohistochemistry – acetone fixed, frozen sections)
Salmeron A, Sanchez-Madrid F, Ursa MA, Fresno M, and Alarcon B. 1991. J. Immunol. 147:3047-3052. (Immunoprecipitation)
Van Dongen JJ, Krissansen GW, Wolvers-Tettero IL, Comans-Bitter WM, Adriaansen HJ, Hooijkaas H, van Wering ER, and Terhorst C. 1988. Blood. 71: 603-612. (Western Blot)
Provided below are standard protocols that you may find useful for product applications.
Background
The UCHT1 antibody is specific for human CD3e, also known as CD3 epsilon, a 20 kDa subunit of the T cell receptor complex, along with CD3 gamma and CD3 delta. These integral membrane protein chains assemble with additional chains of the T cell receptor (TCR), as well as CD3 zeta chain, to form the T cell receptor – CD3 complex. Together with co-receptors CD4 or CD8, the complex serves to recognize antigens bound to MHC molecules on antigen-presenting cells. These interactions promote T cell receptor signaling (T cell activation), inducing cell proliferation, differentiation, production of cytokines or activation-induced cell death. CD3 is differentially expressed during thymocyte-to-T cell development and on all mature T cells.
The UCHT1 antibody is a widely used phenotypic marker for human T cells. In addition, binding/cross-linking of UCHT1 antibody to CD3e can induce cell activation. A recent publication of the crystal structure of a CD3e- antibody complex provides insight as to the action of commonly used agonist antibodies, as well as specific epitope-binding data for the human CD3 antibodies UCHT1 and OKT3 (Fernandes, R.A. et al. 2012. J. Biol. Chem. 287: 13324-13335).
UCHT1 antibody reacts with both surface-expressed and intracellular CD3e protein, in contrast to an alternative human CD3 clone, HIT3a, which will stain only the extracellular (membrane-expressed) CD3e protein. Also, the UCHT1 antibody is reported to be cross-reactive with chimpanzee and has been used for phenotypic analysis of expression by flow cytometry; however the antibody is reported to be unsuitable for induction of T cell activation in this species (Bibollet-Ruche et al. 2009. J. Virol. 82: 10271-10278).
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