0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative. Endotoxin level is <0.1 EU/µg of the protein (<0.01 ng/µg of the protein) as determined by the LAL test.
Preparation
The LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration
1mg/ml
Recommended Usage
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Applications
FC - Quality tested FA, WB, IF - Reported in the literature
Application Notes
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. Additional reported applications (for the relevant formats) include: B cell bioassays. Clone G28.5 has been shown to induce IgG production by B cells7. The G28.5 antibody has been reported to promote B cell proliferation in the presence of anti-IgM, IL-4 or PMA. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm filtered) is recommended for functional assays (Cat. No. 303608). For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 303614) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg).
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Application References
1. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York. 2. Banchereau J, et al. 1994. Annu. Rev. Immunol. 12:881. (FA) 3. Gaspari A, et al. 1996. Eur. J. Immunol. 26:1371. (FA) 4. Ebner S, et al. 2001. J. Immunol. 166:633. (FA) 5. Lin-Lee YC, et al. 2006. J. Biol. Chem. 281:18878. (WB, IF) 6. Jacobsen EM, et al. 2005. Endocrinology 146:2684. (FC) 7. Alderson MR, et al. 1993. J. Exp. Med. 178:669.
Description
CD40 is a 48 kD type I glycoprotein also known as BP50. It is a member of the TNFR superfamily primarily expressed on B cells, macrophages, follicular dendritic cells, endothelial cells, fibroblasts, and at low levels on plasma cells. CD40 has been reported to be involved in B cell differentiation, costimulation, isotype class-switching, and protection from apoptosis. Additionally, CD40 is important for T cell-B cell interactions. The ligand of CD40 is CD154 (CD40 ligand).
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Storage
The antibody solution should be stored undiluted 2°C and 8°C. This LEAF™ solution contains no preservative; handle under aseptic conditions.