Anti-Mouse MAdCAM-1 (MECA-89) – Purified in vivo GOLD^TM Functional Grade

D230

Mouse mesenteric and peripheral lymph node cells

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.

This antibody is stable for at least one week when stored sterile at 2-8°C. For long term storage aseptically aliquot in working volumes without diluting and store at –80°C. Avoid Repeated Freeze Thaw Cycles.

Research Use Only

2 - 8°C Wet Ice

MECA-89 activity is directed against mouse MAdCAM-1.

MAdCAM-1 is a cell adhesion leukocyte receptor expressed by mucosal venules that helps direct lymphocyte traffic into mucosal tissues and regulates the passage and retention of leukocytes^{1, 2}. MAdCAM-1 binds integrin receptor α4β7 and L-selectin^{2, 3, 4}. Two alternatively spliced isoforms of MAdCAM-1 exist⁵, both of which are capable of binding α4β7².

MECA-89 was generated by immunizing Wistar rats with endothelial cells isolated from BALB/c mesenteric and peripheral lymph nodes ⁶. Immunohistological screening of hybridomas yielded two mAbs, MECA-367 and MECA-89, that stain high endothelial venules (HEVs) in mucosal lymphoid organs and Peyer’s patches, but not peripheral lymph nodes (axillary, brachial, popliteal, and inguinal). Immunofluorescence staining of high endothelial cells shows that both MECA-367 and MECA-89 react with antigens on the cell surface. The epitopes for MECA-367 and MECA-89 are distinct. MECA-367 recognizes the N-terminal immunoglobulin domain of MAdCAM-l, and MECA-89 recognizes the second immunoglobulin domain ^4,5.

MECA-89 reacts with the same vessels as MECA-367 and binds to isolated MECA-367 antigen; however, unlike MECA-367 it has no effect on lymphocyte binding ⁶. Additionally, MECA-89 has no effect on MAdCAM-1 binding to α4β7 in vitro ⁷, but L-selectin dependent adhesion is lost in the presence of MECA-89 ⁴. Additionally, MECA-89 has no significant effect on activated cells, but all interactions are inhibited following subsequent injection of anti-α4 Fab fragments ⁴. In vivo, MECA-89 inhibits L-selectin-dependent rolling but not direct α4β7-dependent attachment of Mn2+ activated lymphocytes.

MAdCAM-1 is a cell surface glycoprotein selectively expressed on high endothelial venules of mucosal lymphoid organs and Peyer’s patches as well as lamina propria venules.

Integrin a4ß7, CD62L

1. https://www.uniprot.org/uniprotkb/Q61826/entry
2. Schiffer SG, Day E, Latanision SM, et al. Biochem Biophys Res Commun. 216(1):170-176. 1995.
3. Berlin C, Berg EL, Briskin MJ, et al. Cell. 74(1):185-195. 1993.
4. Bargatze RF, Jutila MA, Butcher EC. Immunity. 3(1):99-108. 1995.
5. Briskin MJ, McEvoy LM, Butcher EC. Nature. 363(6428):461-464. 1993.
6. Streeter PR, Berg EL, Rouse BT, et al. Nature. 331(6151):41-46. 1988.
7. Nakache M, Berg EL, Streeter PR, et al. Nature. 337(6203):179-181. 1989.