CD40 Monoclonal Antibody (5C3), Super Bright™ 645, eBioscience™
Isotype: IgG1, kappa
Format: Super Bright™ 645
Application: Flow Cytometry
Tested Dilution: 5 µL (0.125 µg)/test
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!
Formulation: PBS, pH 7.2, containing 0.09% sodium azide
Purification: Affinity chromatography
Data Sheet: TDS
Description: The 5C3 monoclonal antibody reacts with human CD40, a 45-50 kDa type I transmembrane glycoprotein. CD40 is a member of the TNFR family and is expressed by B lymphocytes, follicular dendritic cells, thymic epithelium, and a subset of peripheral T cells. CD40 regulates B cell development and maturation by inducing Ig isotype-switching and in combination with other signals such as IL-4, protects B cells from surface Ig-induced apoptosis and promotes proliferation. Interaction of CD40 with CD154 (gp39), its ligand on T cells, is important in T-B cell crosstalk and plays a role in costimulation and immune regulation. 5C3 is reported to be used for activation of B cells in in vitro functional assays.
Applications Reported: The 5C3 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 5C3 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at less than or equal to 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Super Bright 645 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 645 nm. We recommend using a 660/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 645 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.