CD40 Human, IgG2a Fc Mouse, Fusion Protein

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. CD40 Human, IgG2a Fc Mouse, 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 (173aa) domain of human CD40 fused to murine IgG2a Fc (233aa).

Applications:
Suitable for use in ELISA and Flow Cytometry. Other applications not tested.

Recommended Dilutions:
Flow Cytometry: 80ul at 5ug/ml labels 5x10e5 Jurkat-4G cells
Optimal dilutions to be determined by the researcher.

Flow Cytometry Protocol:
1. Jurkat-4G (a CD154 positive subclone of Jurkat)cells were stimulated 6 hours in the presence of PMA and Ionomycin (to increase CD154 expression).
2. Five x 10e5 cells per tube were incubated 45 minutes on ice with 80ul of C2392-02K at a concentration of  5ug/ml.
3. Cells were washed twice and incubated with 50ul secondary reagent IgG (FITC) goat anti-mouse, after which they were washed three times, fixed and analyzed by FACS.
4. Cells stained positive with a mean shift of 0.22 log10 fluorescent units when compared to a Mouse IgG2a negative control at a similar concentration.

Storage and Stability:
May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.