Anti-Mouse/Human PNAd (Clone MECA-79) – Purified in vivo GOLD^TM Functional Grade

Mouse lymph node stromal 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 only in vitro 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.

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 -80°C. Avoid Repeated Freeze Thaw Cycles.

Research Use Only

2 – 8° C Wet Ice

MECA-79 reacts with a family of sialomucins known collectively as peripheral node addressin (PNAd). MECA-79 recognizes O-glycans containing 6-sulfo N-acetylglucosamine in the extended core 1 structure.

MECA-79 antibody and L-selectin recognize the same family of sialomucins known collectively as peripheral node addressin (PNAd), with MECA-79 staining ligands for L- selectin expressed by high endothelial venules (HEVs)¹. The MECA-79 epitope consists of a 6-sulfo N-acetyl-lactosamine modification of the extended core-1 branch that overlaps with the sialyl 6-sulfo Lewis X at its terminus. Binding requires GlcNAc-6 sulfation but not sialylation and/or fucosylation. MECA-79 stains lymph node HEVs and blocks L-selectin-dependent lymphocyte attachment in vitro and in vivo in mouse models².

In vitro, MECA-79 blocks normal lymphocytes and a peripheral lymph node-specific lymphoma from binding to peripheral lymph node HEV². In vivo, MECA-79 inhibits normal lymphocyte homing to peripheral lymph nodes. Additionally, MECA-79 detects HEVs in lymph nodes of HEV-like vessels at the sites of chronic inflammation, where PNAd + vessels are induced, and staining of these vessels corresponds to functional L- selectin ligands¹. Furthermore, MECA-79 antibody treatment reduces lymphocyte adhesion to GlyCAM-1 from wildtype mouse HEV³.

MECA-79 was generated by immunizing a Wistar rat with lymphocytes released from pooled axillary, brachial, inguinal, and mesenteric lymph nodes of BALB/c mice². Spleen cells from immune animals were fused with mouse myeloma Sp2/0 to create hybridomas, which were screened by immunofluorescence for an antibody that selectively stains peripheral lymph node HEV. MECA-79 is IgM.

In mice, PNAd staining is detected on all high endothelial venules (HEV) in peripheral lymph nodes with intense cytoplasmic as well as luminal and abluminal cell surface reactivity. MECA-79 stains peripheral lymph nodes, some segments of mesenteric lymph nodes, the abluminal aspect of high endothelial cells (HEC) in Peyer’s patches, and postcapillary venules of inflamed non-lymphoid tissues. In humans, MECA-79 binds to HEC from tonsil.

CD62L

Q64311

1 Uchimura K, Rosen SD. Trends Immunol. 27(12):559-565. 2006.
2 Streeter PR, Rouse BT, Butcher EC. J Cell Biol. 107(5):1853-1862. 1988.
3 Yeh JC, Hiraoka N, Petryniak B, et al. Cell. 105(7):957-969. 2001.
4 Kawashima H, Petryniak B, Hiraoka N, et al. Nat Immunol. 6(11):1096-1104. 2005.
5 Uchimura K, Gauguet JM, Singer MS, et al. Nat Immunol. 6(11):1105-1113. 2005.
6 Hemmerich S, Butcher EC, Rosen SD. J Exp Med. 180(6):2219-2226. 1994.
7 Rosen SD, Tsay D, Singer MS, et al. Am J Pathol. 166(3):935-944. 2005.