hCD70-muCD8 Fusion Protein (human CD70-murine CD8 Fusion Protein)

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