Anti-Mouse CD3 (17A2) – Purified (PhenoCyler-Fusion (CODEX)® Ready)

Clone 17A2 has been validated for use on CODEX® using mouse spleen fresh frozen (FF) tissue.

γ/δ TCR-positive T-T hybridoma D1

This purified antibody is formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.4.

Recombinant antibodies are manufactured in an animal free facility using only <i>in vitro</i> protein free cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.

This antibody is stable for at least one week when stored at 2-8°C. For long term storage, aliquot in working volumes without diluting and store at -20°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.

PhenoCycler-Fusion (CODEX)®
FA^2,3,5,6
Depletion^2,4,5
IP⁶
RIA⁶
FC
IHC (Frozen) The suggested concentration for this 17A2 antibody in IHC staining on frozen tissue is 5.0 - 10 μg per ml. Titration of the reagent is recommended for optimal performance for each application.

AB_2893570

17A2 activity is directed against the mouse TCR ⍺β- and TCR γ𝛿-associated CD3 molecular complex via the CD3ε chain.

CD3 is expressed on T cells.

CD3 is an invariant antigen of the T cell receptor (TCR) ¹. The CD3/TCR complex is composed of a ⍺β or γ𝛿 TCR heterodimer noncovalently associated with invariant CD3 dimers 𝜀γ, 𝜀𝛿, and 𝜁𝜁 in a 1:1:1:1 stoichiometry. The TCR mediates recognition of antigenic peptides bound to major histocompatibility complex (MHC) molecules on antigen-presenting cells, while the CD3 portion of the complex transduces activation signals to the T cell nucleus. Together, TCR and CD3 molecules initiate protective immunity against microbes and cancers.

17A2 has been explored as a treatment against graft-versus-host disease ^2,3. 17A2 leads to T cell depletion in vivo that is associated with a significant reduction in B cell population ⁴. Additionally, 17A2 is less mitogenic that some other anti-CD3 antibodies (i.e., 145-2C11 hamster IgG and KT3 rat IgG2a) ³. FcγRI may be involved in 17A2 activity ⁵.

17A2 was generated by immunizing female Sprague-Dawley rats with homogenized γ𝛿 TCR⁺ T-T hybridoma cells (D1 line) ⁶. The resulting rat cells were fused to P3X63-Ag8-653 non-secreting mouse myeloma cells and hybridomas were screened for the secretion of IL-2 in the presence of D1 cells.

Peptide antigen/MHC-complex

Antigen recognition, TCR signal transduction, T cell activation

1. Mariuzza RA, Agnihotri P, Orban J. J Biol Chem. 295(4):914-925. 2020.
2. Mysliwietz J, Thierfelder S. Blood. 80(10):2661-2667. 1992.
3. Vossen AC, Tibbe GJ, Kroos MJ, et al. Eur J Immunol. 25(6):1492-1496. 1995.
4. Loubaki L, Tremblay T, Bazin R. J Immunol Methods. 393(1-2):38-44. 2013.
5. Kummer U, Zengerle U, Pischel J, et al. Immunol Lett. 75(2):153-158. 2001.
6. Miescher GC, Schreyer M, MacDonald HR. Immunol Lett. 23(2):113-118. 1989.