Detects mouse MAdCAM-1 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human (rh) MAdCAM-1, recombinant mouse (rm) ICAM-1, rmICAM-2, rmICAM-5, rmVCAM-1, and rhICAM-3.
Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse MAdCAM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF993) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for MAdCAM‑1 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
MAdCAM‑1 was detected in perfusion fixed frozen sections of mouse intestine using Goat Anti-Mouse MAdCAM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF993) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the endothelial cells of blood capillaries in intestinal villi. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is an immunoglobulin (Ig) cell adhesion molecule family member. In addition to Ig domains, it contains a mucin-like domain and a membrane proximal domain with similarity to IgA. MAdCAM-1 is involved in lymphocyte homing to mucosal sites and is expressed on high endothelial venules (HEV) of both mesenteric lymph nodes and Peyer’s patches. It has also been found to be expressed on sinus-lining cells of the spleen. The integrin, alpha 4 beta 7, has been shown to function as the MAdCAM-1 receptor. The Ig domains of MAdCAM-1 have been found to be critical to alpha 4 beta 7 binding. The mucin domain has been shown to have activity in L-Selectin binding. MAdCAM-1 expression has been demonstrated to be up-regulated by TNF-alpha and IL-1 beta. MAdCAM-1 appears to play a role in inflammatory bowel disease (IBD) as its expression is highly up-regulated in IBD and most likely serves to recruit alpha 4 beta 7-expressing lymphocytes to the region. In vivo studies involving nonobese diabetic (NOD) mice have also suggested that MAdCAM-1/ alpha 4 beta 7 interaction plays a role in diabetes development in this model. Mouse MAdCAM-1 is a 405 amino acid (aa) residue protein with a 21 aa signal sequence, a 344 aa extracellular domain, a 20 aa transmembrane domain and a 20 aa cytoplasmic domain.
Briskin, M.J. et al. (1993) Nature 363:461.
Yang, X.D. et al. (1997) Diabetes 46:1542.
Sampaio, S.O. et al. (1995) J. Immunol. 155:2477.
Kraal, G. et al. (1995) Am. J. Pathol. 147:763.
Berg, E.L. et al. (1993) Nature 366:695.
Takeuchi, M. and V.R. Baichwal (1995) Proc. Natl. Acad. Sci. USA 92:3561.
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Lymph node tissue,Colon tissue
Other Experimental Details
Other Experimental Details
Tissues fixed in 10% Formalin and embedded in paraffin blocks. Antigen retrieval in Tris-Base EDTA buffer pH 9.0. Incubated overnight with MAdCAM-1 at a concentration of 5ug/ml. Used CTS008 (Anti-Goat HRP-DAB cell and tissue staining kit) and saw signal in endothelial cells with no background.