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Invitrogen™ CD40 Monoclonal Antibody (5C3), eFluor™ 450, eBioscience™

Description
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: This 5C3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 5C3 antibody has been pre-titrated and tested by flow cytometric analyis of normal human peripheral blood cells. This can be used at 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. eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser. Filtration: 0.2...
Specifications
Specifications
| Antigen | CD40 |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | 5C3 |
| Concentration | 5 μL/Test |
| Conjugate | eFluor 450 |
| Formulation | PBS with BSA and 0.09% sodium azide; pH 7.2 |
| Gene | CD40 |
| Gene Accession No. | P25942 |
| Gene Alias | AI326936; B cell surface antigen CD40; B cell-associated molecule; B-cell surface antigen CD40; Bp50; CD antigen CD40; Cd40; CD40 antigen; CD40 antigen (TNF receptor superfamily member 5); CD40 antigen, TNF receptor superfamily member 5; CD40 molecule; CD40 molecule, TNF receptor superfamily member 5; CD40L receptor; CDw40; GP39; HIGM1; I79_006806; IGM; IMD3; Immunoglobulin M; ImmunoglobulinM; membrane protein CD40; MGC9013; p50; receptor for ligand CD154; sCD40; soluble CD40; T-BAM; T-cell differentiation antigen; Tnfrsf5; TRAP; tumor necrosis factor receptor superfamily member 5; tumor necrosis factor receptor superfamily, member 5 |
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Frequently Asked Questions (FAQs)
Our options will depend on the samples you are analyzing.
If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.
For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.
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