Anti-CD152, Blocking (CTLA4, Cytolytic T-lymphocyte-associated Antigen 4), Biotin conjugated, clone

Human CD152 (CTLA-4) is a cell surface glycoprotein expressed at low levels on activated T-cells. CD152 is a high affinity receptor for the costimulatory molecules CD80 (B7-1) and CD86 (B7-2) and appears to function as a negative regulator of T cell activation. Applications: Suitable for use in Flow Cytometry, ELISA, Blocking, Immunocytochemistry and Immunohistochemistry. Other applications not tested. Recommended Dilution: Immunohistochemistry: Frozen sections, ELISA: 1ng/ml, Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Ficoll prepared human peripheral blood lymphocytes were stimulated in culture 3 days in the presence of PHA (5ug/ml). 2. 5x10e5 cells were washed and pre-incubated five minutes on ice with 20ul of 250ug/ml human Ig (to block nonspecific binding) and then incubated 45 minutes on ice with 80ul of C2548-83C1 at 20ug/ml. , 3. Cells were washed twice and incubated with secondary reagent S7973-88P, after which they were washed three times, fixed and analyzed using a BD FACstar plus. , 4. The cells stained positive with a mean shift of 2.28 log10 fluorescent units when compared to Mouse IgG1/Biotin negative control. , 5. Binding was 99% blocked when cells were first pre incubated 10 minutes with 20ul of 0.5mg/ml C2548-83C antibody. Hybridoma: NS1 myeloma cell line , Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.