Invitrogen

CD40 Monoclonal Antibody (5C3), NovaFluor™ Yellow 755, eBioscience™

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Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

H027T03Y08-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.6 µg)/test
-
Product Specifications

Species Reactivity

Human

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

5C3

Conjugate

NovaFluor™ Yellow 755 NovaFluor™ Yellow 755 NovaFluor™ Yellow 755

Excitation/Emission Max

551/755 nm View spectra spectra

Form

Liquid

Concentration

0.6 µg/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3098353 (AB_2925878)

Product Specific Information

Description
The 5C3 monoclonal antibody reacts with human CD40, a 45-50 kDa type I transmembrane glycoprotein. 5C3 is reported to be used for activation of B cells in in vitro functional assays.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

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.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• 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.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 551 nm; Emission: 755 nm; Laser: 561 nm (Yellow) 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.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

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Bioinformatics

Protein Aliases: B cell surface antigen CD40; B cell-associated molecule; B-cell surface antigen CD40; Bp50; CD antigen CD40; CD40; CD40 molecule, TNF receptor superfamily member 5; CD40L receptor; CDw40; Immunoglobulin M; ImmunoglobulinM; MGC9013; sCD40; soluble CD40; Tumor necrosis factor receptor superfamily member 5

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Gene Aliases: Bp50; CD40; CDW40; p50; TNFRSF5

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UniProt ID: (Human) P25942

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Entrez Gene ID: (Human) 958

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