Human CD51 is an integrin alpha V chain which can associate with five different integrin beta chains to form various adhesion molecules. The alpha V beta 3 integrin complex binds vitronectin at the RGD sequence and also can bind to fibrinogen, von Willebrand factor, thrombospondin, fibronectin, osteopontin and collagen. Applications: Suitable for use in Flow Cytometry. Other applications not tested. Recommended Dilution: Flow Cytomertry: 10ug/ml. Use 80ul to label 5x10e5 cultured UM-SCC cells. Immunohistochemistry: Frozen Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Five x 10e5 cultured UM-SCC (squamous cell carcinoma) cells were harvested by trypsinization, washed and preincubated for 5 minutes with 20ul of 250ug/ml human IgG (To block nonspecific binding), after which they were incubated for 45 minutes on ice with 80ul of C2405-13B at 10ug/ml. 2. Cells were washed twice and incubated with secondary reagent Streptavidin (PE), S7973-88P, after which they were washed three times, fixed and analyzed by FACS. 3. Cells stained positive with a mean shift of 2.05 log10 fluorescent units when compared to a Mouse IgG1/Biotin negative control at a similar concentration. 4. Binding was blocked when pre-incubated for 10 minutes with 20ul of 0.5mg/ml C2405-13A, CD51 antibody. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
