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Invitrogen
CD40 Monoclonal Antibody (5C3), NovaFluor™ Red 755, eBioscience™
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FIGURE: 1 / 2
CD40 Antibody (H027T02R06) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD40 Monoclonal Antibody, NovaFluor Red 755 (Product # H027T03R06) (right). All cells were co-stained with CD19 Monoclonal Antibody, eFluor 450 (Product # 48-0199-42). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
H027T02R06
Product Specifications
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
5C3
Conjugate
Excitation/Emission Max
636/755 nm
View spectra
Form
Liquid
Concentration
0.6 µg/Test
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Product Specific Information
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: The 5C3 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at less than or equal to 5 µL (0.6 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.
NovaFluor dyes are not compatible with DNA intercalating viability dyes. Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100µL of cell sample containing 10^3 to 10^8 cells.
NovaFluor conjugates are based on Phiton™ technology utilizing novel nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 636 nm; Emission: 756 nm; Laser: 633-640 nm (Red) Laser
Target Information
CD40 is a single-chain glycoprotein and a member of the tumor necrosis factor receptor (TNFR) family, exhibiting significant homology to the Hodgkin's disease-associated antigen, CD30. It is expressed by B lymphocytes, follicular dendritic cells, thymic epithelium, and a subset of peripheral T cells, as well as some epithelial cells, carcinomas, and lymphoid dendritic cells. Notably, CD40 is present on all B cells except plasma cells. CD40 plays a crucial role in regulating B cell development and maturation, inducing immunoglobulin isotype-switching, and protecting B cells from surface Ig-induced apoptosis when combined with other signals such as IL-4. It promotes proliferation and is essential for T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. The interaction between CD40 and its ligand CD154 (gp39) on T cells is vital for T-B cell crosstalk, costimulation, and immune regulation. Adaptor protein TNFR2 interacts with CD40, mediating signal transduction, while the AT-hook transcription factor AKNA is reported to regulate CD40 expression, which may be important for homotypic cell interactions. Additionally, the interaction between CD40 and its ligand is necessary for amyloid-beta-induced microglial activation, suggesting a role in the early pathogenesis of Alzheimer's disease. Two alternatively spliced transcript variants encoding distinct isoforms of CD40 have been identified. Diseases associated with CD40 dysfunction include Type 3 Hyper-IgM immunodeficiency and CD40 ligand deficiency, highlighting its importance in immune and inflammatory responses.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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