Western Blotting (WB), Flow Cytometry (FACS), Enzyme Immunoassay (EIA)
Specificity
This antibody recognises Mucosal Addressin Cell Adhesion Molecule 1 (MAdCAM-1). MAdCAM-1 expression has also been reported in the gut lamina propria but Clone 314G8 does not recognise MAdCAM-1 in these tissues. Clone 314G8 reacts with the ligand-binding first Ig domain and reports suggest that a splice variant exists in the gut which is not recognised by clone 314G8. Clone 314G8 is reported to block the interaction of MAdCAM-1 with alpha 4 beta 7. We recommend the use of SM2237LE for this purpose.
Cross-Reactivity (Details)
Species reactivity (tested):Human.
Purification
Affinity Chromatography on Protein G.
Immunogen
Recombinant soluble MAdCAM-1-fc fusion protein. Spleen cells from immunised Balb/c mice were fused with cells of the mouse P3X63Ag8.653 myeloma cell line.
ELISA. Western Blot: under reducing conditions, This antibody detects a band of approximately 60 kDa. Flow Cytometry: 1/10- 1/100, use 10 μL of the suggested working dilution to label 10^6 cells in 100 μL. Immunohistochemistry on paraffin embedded sections: 1/50, This product does not require protein digestion pre-treatment of paraffin embedded sections prior to staining. This product does not require antigen retrieval prior to staining of paraffin sections. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Concentration
1.0 mg/mL
Buffer
PBS, 0.09 % Sodium Azide
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C/-20 °C
Storage Comment
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Olloquequi, Montes, Prats, Rodríguez, Montero, García-Valero, Ferrer: "Significant increase of CD57+ cells in pulmonary lymphoid follicles of COPD patients." in: The European respiratory journal, Vol. 37, Issue 2, pp. 289-98, (2011) (PubMed).
Leung, Lehnert, Kanwar, Yang, Mon, McNeil, Krissansen: "Bioassay detects soluble MAdCAM-1 in body fluids." in: Immunology and cell biology, Vol. 82, Issue 4, pp. 400-9, (2004) (PubMed).
MAdCAM-1 is a 60kD cell surface protein that is involved in lymphocyte trafficking. MAdCAM-1 is expressed on high endothelial venules of Peyers patches and mesenteric lymph nodes. MAdCAM-1 expression has also been reported in the gut lamina propria.Synonyms: MAdCAM-1, Mucosal addressin cell adhesion molecule 1, hMAdCAM-1