Catalogue

Background
Integrins are transmembrane glycoproteins that belong to the family of adhesion molecules. They promote interactions between cells and their environment, being both other cells or the extra- cellular matrix. Integrins are a family of heterodimeric membrane glycoproteins consisting of non-covalently associated subunits, i.e. an α-subunit of 95 kDa that is conserved through the superfamily and a more variable β subunit of 150-170 kDa. More than 18 α and 8 β subunits with numerous splice variant isoforms have been identified in mammals. The integrin α5 chain (160 kDa) (CD49e), undergoes post-translational cleavage in the extracellular domain to yield disulfide-linked light (25 kDa) and heavy (135 kDa) chains, which non-covalently associate with the integrin β1 subunit (CD29) (130 kDa), thus forming the heterodimer α5-β1 very late antigen (VLA-5) complex. VLA-5 is a fibronectin receptor (FNr) that is expressed on thymocytes, T-cells, monocytes and platelets. It is also found on very early B-cells and activated B-cells. VLA-5 mediates binding of T- and B-cells to fibronectin, an extracellular matrix glycoprotein involved in cell adhesion and migRation in wound healing, embryonic development and malignant transformation. The VLA-5 or FNr binds only fibronectin recognizing the Arg-Gly-Asp (RGD) sequence in the central region of the molecule.

Source
NKI-SAM-1 is a monoclonal antibody obtained by fusion of mouse myeloma cells with spleen cells from BALB/C mice immunized with U937 histiocytic lymphoma cells.

Product
Each vial contains 100 ul 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Formulation: Each vial contains 100 ul 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Specificity
NKI-SAM-1 recognizes the 135-kDa integrin- alpha 5 monocyte-specific antigen. As a result NKI-SAM-1 strongly reacts with monocytes, and weakly with granulocytes, platelets and T-lymphocytes. Its exact epitope is not known.

Applications
NKI-SAM-1 is suitable for flow cytometry (recommended range is 1:200 – 1:500, using 0.2-0.5 µg for 10^6 cells) and can be used to determine the VLA-5 expression in peripheral blood monocytes and monocyte derived dendritic cells (see refs 2 and 3), B cells and B-cell precursors (ref. 4), and in the less pathogenic yeast Candida species (ref. 5). NKI-SAM-1 can also be applied in immunohisto-chemistry on frozen tissues, immunocytochemistry and immunoprecipitation. Optimal antibody dilution should be determined by titration.

Storage
The antibody is shipped at ambient temperature and may be stored at +4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.

Shipping Conditions: Ship at ambient temperature

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but our company accepts no liability for any inaccuracies or omissions in this information.

References
1. Te Velde, A., Klomp, J., Yard, B., De Vries, J., Figdor, C. (1988). Modulation of phenotypic and functional properties of human peripheral blood monocytes by IL-4. J Immunol 140, 1548-54. 2. 2. Ammon, C., Meyer, S.P., Schwarzfischer, L., Krause, S.W., Andreesen, R., Kreutz, M. (2000). Comparitive analysis of integrin expression on monocyte-derived macrophages and monocyte-derived dendritic cells. Immunology 100, 364-9. 3. 3. Grassi, F., Dezutter-Dambuyant, C., McIlroy, D., Jacquet, C., Yoneda, K., Imamura, S., Boumsell, L., Schmitt, D., Autran, B., Debre, P., Hosmalin, A. (1998). Monocyte-derived dendritic cells have a phenotype comparable to that of dermal cells and display ultrastructural granules distinct from Birbeck granules. J Leukoc Biol 64, 484-93. 4. 4. Xiao, J., Messinger, Y., Jin, J., Myers, D., Bolen, J., Uckun, F. (1996). Signal transduction through the b1 integrin family surface adhesion molecules VLA-4 and VLA-5 of human B-cell precursors activates CD19 receptor-associated protein-tyrosine kinases. J Biol Chem 271, 7659-64. 5. 5. Santoni, G., Birbarelli, P., Hong, L., Gamero, A., Djeu, J., Piccoli, M. (1995). An ?5?1-like integrin receptor mediates the binding of less pathogenic Candida species to fibronectin. J Med Microbiol 43, 360-7. 6. 6. Klingemann, H.G., Dedhar, S. (1989). Distribution of integrins on human peripheral blood mononuclear cells. Blood 74, 1348-54.

Safety Datasheet(s) for this product: