The antibody was purified by affinity chromatography, and conjugated with FITC under optimal conditions. The solution is free of unconjugated FITC.
Recommended Usage
Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. Test size products are transitioning from 20 µl to 5 µl per test. Please check your vial or your CoA to find the suggested use of this reagent per million cells in 100 µl staining volume or per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application. Read more at www.biolegend.com/testsize regarding the test size change.
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Applications
ICFC - Quality tested
Application Notes
ELISA Capture1 or ELISPOT Capture2: The purified DB-1 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated poly5109 antibody (Cat. No. 510901) as the detecting antibody and recombinant IFN-γ (Cat. No. 565701) as the standard. The LEAF™ purified antibody is suggested for ELISPOT capture. Flow Cytometry5: The fluorochrome-labeled DB-1 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IFN-γ-producing cells within mixed cell populations. For intracellular cytokine staining protocol, please visit www.biolegend.com and click on the support section. Neutralization3,4: The LEAF™ purified antibody (Endotoxin in vivo and in vitro (Cat. No. 507808). Additional reported applications (for the relevant formats) include: Western blotting1, and immunohistochemistry2 of paraformaldehyde-fixed, saponin-treated frozen tissue sections.
Application References
1. Van der Meide P, et al. 1989. Lymphokine Res. 8:439. 2. Nennesmo I, et al. 1989. Brain Res. 504:306. 3. Rayner D, et al. 1987. Scand. J. Immunol. 25:621. 4. Hartung H, et al. 1990. Ann Neurol. 27:247. 5. Bernard I, et al. 1998. Eur. Cytokine Net. 9:613. 6. Nakagawa P, et al. 2012. Am J Physiol Heart Circ Physiol. 303:1114. PubMed.
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Description
Interferon-γ is a potent multifunctional cytokine which is secreted primarily by activated NK cells and T cells. Originally characterized based on anti-viral activities, IFN-γ also exerts anti-proliferative, immunoregulatory, and proinflammatory activities. IFN-γ can upregulate MHC class I and II antigen expression by antigen-presenting cells. The DB-1 antibody reacts with rat and mouse interferon-gamma (IFN-γ). The DB-1 antibody can neutralize the bioactivity of natural or recombinant IFN-γ. The DB-1 antibody has been well characterized for ELISPOT, ELISA, intracellular staining, Western blotting, IHC, and neutralization (in vitro and in vivo).
Storage
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.