CD40 Human, IgG2a Fc Mouse, Fusion Protein (PE)

Human CD40 is a member of the tumor necrosis factor (TNF) receptor family and is present on all B cells except plasma cells. CD40 is also found on some epithelial cells, carcinomas and lymphoid dendritic cells. CD40 plays an important role in B cell activation and the interaction with its ligand CD154 (CD40 Ligand) is essential for isotype switching (1). CD40 muIg fusion protein is recognized by anti-human CD40 antibody and binds to human CD154. This binding can be blocked by anti-CD154 antibody.||Molecular Structure: |A soluble fusion protein consisting of the extracellular (193aa) domain of human CD40 fused to murine IgG2a Fc (232aa), with a predicted monomeric molecular weight of 45.6kD.||Applications: |Suitable for use in ELISA and Flow Cytometry. Other applications not tested.||Functional Application: |CD40-muIg fusion protein is recognized by anti-human CD40 (C2392-02E). This binding can be blocked by using anti-human CD154 (C2548-84W1).||Recommended Dilutions:|Flow Cytometry: 1:20 (20ug/ml). 80ul of diluted solution labels 5x10e5 cells.|Optimal dilutions to be determined by the researcher.||Storage and Stability:|Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 6 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: PE conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.||Flow Cytometry Protocol:|1. Ficoll prepared human peripheral blood mononuclear cells were stimulated 6 hours in the presence of 10ng/ml PMA and 1uM Ionomicin to upregulate CD154 expression. |2. Five x 10e5 cells per tube were washed and incubated for 45 minutes on ice with 80ul of a 1:20 dilution (20ug/ml) of C2392-02L. |3. They were washed three times, fixed and analyzed by Flow Cytometry. |4. A 39% sub-population of the cells stained positive with a mean shift of 1.16 log10 fluorescent units when compared to a negative control.|5. Binding was effectively blocked when cells were preincubated 10 minutes with 20ul of 0.5mg/ml anti-CD154 antibody (C2548-84W1).