Description: The 1C10 monoclonal antibody reacts with mouse CD40, a 45-50 kDa type I transmembrane glycoprotein. CD40 is a member of the TNFR family and is expressed by mouse B lymphocytes, follicular dendritic cells, thymic epithelium, and a subset of peripheral T cells. CD40 regulates B cell development/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. The monoclonal antibody 1C10 is reported to have agonistic activity in vitro and in vivo. Applications Reported: The 1C10 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 1C10 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 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. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5.We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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: 488 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered. The CD40 antigen is a single chain glycoprotein that is known to be a member of the tumor necrosis factor/nerve growth factor superfamily and shows a significant homology to the Hodgkin ft.s disease associated antigen, CD30. CD40 is present on all B cells except plasma cells and is also found on some epithelial cells, carcinomas and lymphoid dendritic cells. CD40 has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. AT-hook transcription factor AKNA is reported to coordinately regulate the expression of CD40, which may be important for homotypic cell interactions. Adaptor protein TNFR2 interacts with CD40 and serves as a mediator of the signal transduction. The interaction of CD40 and its ligand is found to be necessary for amyloid-beta-induced microglial activation, and thus is thought to be an early event in Alzheimer disease pathogenesis. Two alternatively spliced transcript variants of CD40 encoding distinct isoforms have been reported. Diseases associated with CD40 dysfunction include Type 3 Hyper-Igm immunodeficiency and CD40 ligand deficiency.
|Store at 2-8°C. Do not freeze. Light-sensitive material.|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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