IFN-α 抑制的体外试验

Kathrin Gibbert

doi:10.21769/BioProtoc.802

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Peer-reviewed

IFN-α Inhibition Assay in vitro

IFN-α 抑制的体外试验

Kathrin Gibbert email

发布: 2013年06月20日第3卷第12期 DOI: 10.21769/BioProtoc.802 浏览次数: 10363

评审: Fanglian HeLin Fang

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参见作者原研究论文

The authors used this protocol in:

Cover of PLOS Pathogens, featuring study using the protocol.

Aug 2012

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Abstract

During viral infections Interferon-α (IFN-α) is expressed by infected host cells. IFN-α binds to its receptor (IFNAR1/2), which leads to the activation of downstream signaling via JAK-STAT. This signaling cascade results in the expression of several hundred different genes, so called interferon-stimulated gene, which lead to an antiviral state of the infected and the neighboring cells.

Keywords: Type I IFNs (I型IFNs)

IFN-alpha subtypes (IFN-α亚)

Friend Murine leukemia virus (Friend Murine leukemia病毒)

Antiviral effect (抗病毒作用)

Materials and Reagents

Mus dunni fibroblast cells
Friend murine leukemia virus (F-MuLV) (titer was measured as previously described (Robertson et al., 1991))
3-Amino-9-ethylcarbazole (AEC) (Sigma-Aldrich, catalog number: A6926-100TAB )
Antibody 720 (mouse antibody against FV envelope protein), hybridoma supernatant (Robertson et al., 1991)
Bovine serum albumin (BSA) (PAA Laboratories GmbH, catalog number: K41-001 )
Ethanol 96% (Roth North America, catalog number: 5054.5 )
Superior FBS (fetal bovine serum, not heat-inactivated) (Biochrom, catalog number: S0615 )
Goat anti-mouse HRP (Dako, catalog number: P0477 )
Hydrogen peroxide 30% (Applichem, catalog number: A1134,0250)
N,N-dimethylformamide (Merck Millipore, catalog number: 1.03053.1000 )
PBS (GIBCO, catalog number: 14190-136 )
Penicillin/streptomycin (PAA Laboratories GmbH, catalog number: P11-010 )
Polybrene/Hexadimethrine bromide (Sigma-Aldrich, catalog number: H9268 )
RPMI 1640 (PAA Laboratories GmbH, catalog number: E15-840 )
Sodium Acetate (Merck Millipore, catalog number: 1062811000 )
Murine IFN-α (PBL, catalog number: 12100-1 )
Medium (see Recipes)
Washing Buffer (see Recipes)
3-Amino-9-ethylcarbazole (AEC) substrate solution (see Recipes)

Equipment

Incubator (37 °C; 5% CO₂)
24 well cell culture plate (Greiner Bio-one, catalog number: 662160 )

Procedure

English

中文翻译

文章信息

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如何引用

Readers should cite both the Bio-protocol article and the original research article where this protocol was used:

Gibbert, K. (2013). IFN-α Inhibition Assay in vitro. Bio-protocol 3(12): e802. DOI: 10.21769/BioProtoc.802.
Gibbert, K., Joedicke, J. J., Meryk, A., Trilling, M., Francois, S., Duppach, J., Kraft, A., Lang, K. S. and Dittmer, U. (2012). Interferon-alpha subtype 11 activates NK cells and enables control of retroviral infection. PLoS Pathog 8(8): e1002868.