Anti-mouse/rat MHC Class II (I-Ek/RT1-D) (Clone 14-4-4S (HB-32)) – Purified in vivo PLATINUM™ Functional Grade

C3H mouse skin graft and splenocytes

This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.

Liquid

Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro 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.

To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM^TM antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.

Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.

Research Use Only

2 – 8° C Wet Ice

14-4-4S (HB-32) activity is directed against mouse MHC Class II alloantigen I-Ek and rat MHC Class II alloantigen RT1-D. In mouse, 14-4-4S cross-reacts with H-2k, H-2d, H-2p, and H-2r and is specific to Ia.7 in the Ia chart. 14-4-4S also recognizes a cell surface marker on hamster B lymphocytes.

H-2, the murine major histocompatibility complex (MHC), is composed of a diverse group of antigens divided into class I and II proteins that function in immune response¹. Class II molecules, also known as Ia antigens, regulate recognition of foreign antigens on the surfaces of antigen presenting cells and play a major role in the mixed lymphocyte response². Mice have two class II isotypes, I-A and I-E, each of which is a glycoprotein composed of an ⍺ and β Subunit.

14-4-4S was generated by immunizing C3H.SW mice with spleen cells from C3H mice³. The resulting spleen cells were then fused with Sp2/0-Ag14 myeloma cells for hybridoma generation. Further studies have found that whole 14-4-4S antibody and F(ab’)₂ fragments block RT1-D-restricted Ag presentation as well as activation of an I-Eʹ-restricted T cell hybridoma in vitro⁴.

Additionally, because 14-4-4S recognizes hamster B cells, 14-4-4S can be used in hamster to deplete B cells and enrich for T cells via flow cytometry⁵ or by removing adherent B lymphocytes in immunoglobulin-coated tissue culture dishes⁶.

I-Ek and RT1-D are expressed primarily on the surface of B lymphocytes, antigen-presenting cells, macrophages, dendritic cells, and a subset of T cells.

CD3/TCR, CD4

1 Yoshida R. Adv Immunol. 124:207-247. 2014.
2 Spencer JS, Kubo RT. J Exp Med. 169(3):625-460. 1989.
3 Ozato K, Mayer N, Sachs DH. J Immunol. 124(2):533-540. 1980.
4 Haag S, Tuncel J, Thordardottir S, et al. J Immunol. 194(6):2539-2350. 2015.
5 Liu H, Steiner BM, Alder JD, et al. Infect Immun. 58(6):1685-1690. 1990.
6 Lim LC, England DM, DuChateau BK, et al. Infect Immun. 63(4):1400-1408. 1995.
7 Shih FF, Racz J, Allen PM. J Immunol. 176(6):3438-3448. 2006.
8 Epstein SL, Ozato K, Bluestone JA, et al. J Exp Med. 154(2):397-409. 1981.
9 Turkewitz AP, Sullivan CP, Mescher MF. Mol Immunol. 20(11):1139-1147. 1983.
10 Maloy WL, Ozato K, Sachs DH, et al. Mol Immunol. 23(3):263-269. 1986.
11 Farr A, DeRoos PC, Eastman S, et al. Eur J Immunol. 26(12):3185-3193. 1996.
12 Desmet C, Gosset P, Pajak B, et al. J Immunol. 173(9):5766-5775. 2004.
13 Holmberg D, Forsgren S, Forni L, et al. Proc Natl Acad Sci U S A. 81(10):3175-3179. 1984.
14 Isobe M, Narula J, Southern JF, et al. Circulation. 85(2):738-746. 1992.