*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The MD-1 antibody reacts with human, rhesus, cynomolgus and baboon interferon-gamma (IFN-γ), the signature Th1 cytokine, which is produced mainly by activated T cells, NK cells and B cells. IFN-γ is a dimeric protein. IFN-γ has antiviral and antiparasitic activities and also inhibits the proliferation of normal and transformed cells.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The MD-1 antibody has been reported for use in intracellular flow cytometric analysis, cytokine neutralization, immunohistochemical staining, ELISA, and ELISPOT. (Fluorochrome conjugated MD-1 is recommended for use in intracellular flow cytometry.) (Please use Functional Grade purified MD-1, cat. 16-7317, in functional assays.)
Applications Tested
The MD-1 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of human interferon-γ (IFN-γ) in combination with the biotinylated 4S.B3 (13-7319) antibody for detection and recombinant human IFN-γ (39-8319) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/ml - 8 pg/ml should be included in each ELISA plate. The FG Purified format of the MD-1 antibody has been tested as the capture antibody in an ELISPOT assay (in conjunction with the biotinylated 4S.B3 antibody) and for neutralization of IFN-γ biological activity. A suitable range of concentrations of this antibody for ELISPOT capture is 1-4 µg/ml. For in vitro neutralization, the MD-1 antibody at 30 ug/ml has been found to neutralize by 50% the effect of 1 ng/ml human IFN-γ. A more potent neutralizing antibody is clone NIB42.
References
Gong G, Shao L, Wang Y, Chen CY, Huang D, Yao S, Zhan X, Sicard H, Wang R, Chen ZW. Phosphoantigen-activated V gamma 2V delta 2 T cells antagonize IL-2-induced CD4+CD25+Foxp3+ T regulatory cells in mycobacterial infection. Blood. 2009 Jan 22;113(4):837-45. (MD-1, cross to cynomolgus, NU, PubMed)
Jurado JO, Alvarez IB, Pasquinelli V, Martínez GJ, Quiroga MF, Abbate E, Musella RM, Chuluyan HE, García VE. Programmed death (PD)-1:PD-ligand 1/PD-ligand 2 pathway inhibits T cell effector functions during human tuberculosis. J Immunol. 2008 Jul 1;181(1):116-25. (MD-1, FA, PubMed)
van der Meide PH, Groenestein RJ, de Labie MC, Heeney J, Pala P, Slaoui M. Enumeration of lymphokine-secreting cells as a quantitative measure for cellular immune responses in rhesus macaques. J Med Primatol. 1995 Dec;24(4):271-81.
Wassenaar A, Reinhardus C, Thepen T, Abraham-Inpijn L, Kievits F. Cloning, characterization, and antigen specificity of T-lymphocyte subsets extracted from gingival tissue of chronic adult periodontitis patients. Infect Immun. 1995 Jun;63(6):2147-53. (MD-1, IHC frozen)
van Besouw NM, van der Meide PH, Bakker NP. The mitogen-induced generation of interferon-gamma producing cells in cultures of rhesus monkey peripheral blood mononuclear cells is age-dependent. J Med Primatol. 1994 Jan;23(1):42-8.
Yazdanbakhsh M, Sartono E, Kruize YC, Kurniawan A, van der Pouw-Kraan T, van der Meide PH, Selkirk ME, Partono F, Hintzen RQ, van Lier RA, et al. Elevated levels of T cell activation antigen CD27 and increased interleukin-4 production in human lymphatic filariasis. Eur J Immunol. 1993 Dec;23(12):3312-7.
Van der Meide PH, Dubbeld M, Schellekens H. Monoclonal antibodies to human immune interferon and their use in a sensitive solid-phase ELISA. J Immunol Methods. 1985 May 23;79(2):293-305.
