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Each lot of this antibody is quality control tested by ELISA analysis. For ELISA capture applications, the suggested use of this reagent is 1-4 ug/ml. To obtain a linear standard curve, serial dilutions of IFN-γ recombinant protein ranging from 1000 to 8 pg/ml are recommended for each ELISA plate. The purified DB-1 has been tested by blocking fluorochrome conjugated DB-1 for intracellular cytokine staining. In order to obtain complete blocking results, a saturated amount of purified antibody (≤ 5.0 ug/million cells) should be used for incubation with target cells, prior to staining with fluorochrome conjugated antibody. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
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. View intracellular cytokine staining protocol Neutralization3,4: The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of rat IFN-γ bioactivity 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.
PMA/lonomycin stimulated Lou rat splenocytes were stained with DB-1 PE.
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).
Other Names:
Interferon-γ, Immune interferon, Type II interferon, T cell interferon, Macrophage-activating factor (MAF), IFN-g, IFN-gamma
Structure:
Cytokine; dimer; 40-80 kD (Mammalian)
Regulation:
Upregulated by IL-2, bFGF, EGF; downregulated by 1-α-25-Dihydroxy vitamin D3, dexamethasone
Cellular Sources:
CD8+ and CD4+ T cells, NK cells
Cellular Targets:
T cells, B cells, macrophages, NK cells, endothelial cells, fibroblasts
Receptors:
IFN-γRα (CDw119) dimerized with IFN-γRβ (AF-1)
Bioactivity/Activities:
Antiviral/antiparasitic activities; inhibits proliferation; enhances MHC class I and II expression on APC
Antigen References:
1. Fitzgerald K, et al. Eds. 2001. The Cytokine FactsBook. Academic Press San Diego. 2. De Maeyer E, et al. 1992. Curr. Opin. Immunol. 4:321. 3. Farrar M, et al. 1993. Annu .Rev. Immunol. 11:571. 4. Gray P, et al. 1987. Lymphokines 13:151.
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This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
FITC anti-rat IFN-γ
PMA/lonomycin stimulated Lou rat splenocytes were stained with DB-1 PE.
LEAF™ Purified anti-rat IFN-γ
PMA/lonomycin stimulated Lou rat splenocytes were stained with DB-1 PE.
PE anti-rat IFN-γ
PMA/lonomycin stimulated Lou rat splenocytes were stained with DB-1 PE.
Purified anti-rat IFN-γ
PMA/lonomycin stimulated Lou rat splenocytes were stained with DB-1 PE.
Alexa Fluor® 647 anti-rat IFN-γ
PMA+ionomycin-stimulated Lou rat splenocytes (6 hours) stained with DB-1 Alexa Fluor® 647
