Human CD178 (FAS ligand CD95L) is a 40kD type II membrane protein that induces apoptosis in cells expressing CD95 (FAS) (1, 2). CD178 is one of the major effectors of CD8+CTL and NK activitity. Loss of CD178 leads to lymphoproliferative and autoimmune disease.
Suitable for use in ELISA, Flow Cytometry and Immunofluorescence. Other applications not tested.
Flow Cytometry: 1:50 (or 1.6ul of concentrated stock/5x10e5 cells)
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Five x 10e5 cultured KFL9 cells (full length CD178 transfectants) per tube were washed and preincubated for 5 minutes with 20ul of 250ug/ml human IgG (to block non-specific binding).
2. Cells were then incubated 45 minutes on ice with 80ul of C2548-91F at a dilution of 1:50 (20ug/ml).
3. Cells washed three times, fixed and analyzed by FACS.
4. Cells stained positive with a mean shift of 0.26 log10 fluorescent units when compared to a mouse IgG1/FITC negative control (I1904-78N) at a similar concentration.
5. Binding was blocked when cells were pre-incubated with 20ul of C2548-91D at 0.5mg/ml.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. 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. FITC conjugates are sensitive to light.