CD70 IgG2a Fc, Fusion Protein (Tnfsf7) (Preservative Free)

Human CD70 (TNFSF7) is the ligand for CD27 (TNFRSF7) and is notably absent on resting lymphocytes. CD70 is expressed after activation on about 20% of T cells and on about 70% of B cells. Recombinant CD70-muCD8 binds to cell surface CD27 on human peripheral blood leukocytes. Molecular Structure:  A soluble molecule consisting of the extracellular domain (167aa) of murine CD8 alpha fused to 146 (c-term) amino acids of the extracellular domain of human CD70, with a predicted monomeric weight of 34.8kD. Extracellular mature murine CD8alpha: (28)KPQAPEL....GLDFACD(194), Linking amino acids: GT, extracellular human CD70: (48)LPLESL....VQWVRP(193). Applications: Suitable for use in ELISA and Flow Cytometry. Other applications not tested. Recommended Dilutions: Flow Cytometry: 10ug/ml, 80ul labels 5x10e5 ficoll prepared human peripheral blood mononuclear cells., Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. 5x10e5 ficoll prepared human peripheral blood mononuclear cells were washed and pre incubated for 5 minutes with 20ul of 250ug/ml human IgG (to block non specific binding)., 2. They were incubated for 45 minutes on ice with 80ul of C2425-01QX at 10ug/ml. 3. Cells were washed twice and incubated with C2259-11D, after which they were washed three times, fixed and analyzed by FACS. , 4. A net 57% sub population of the cells stained positive with a mean shift of 1.15 log10 fluorescent units when compared to a buffer control. Binding was 99% blocked when reagent was first pre incubated with an excess of CD-70 antibody. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer