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Universal type 1 ifn alpha

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The Universal Type I IFN Alpha is a laboratory product used to detect and measure type I interferon alpha in various biological samples. It serves as a standardized reagent for assessing the presence and levels of this important signaling protein.

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Lab products found in correlation

Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Z aad cmk, by Enzo Life Sciences (1 mentions) Cgamp, by InvivoGen (1 mentions) Ruxolitinib, by LC Laboratories (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Prl tk, by Promega (1 mentions) Dual luciferase reporter assay system, by Promega (1 mentions) Lipofectamine 2000, by PBL Assay Science (1 mentions) Facscanto 2, by BD (1 mentions) Versene, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Universal Type I IFN (16 citations) IFN-I (5 citations) Type I IFN (2 citations) Universal IFN-I (2 citations)

5 protocols using universal type 1 ifn alpha

1

Interferon Alpha Signaling Modulation

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Universal Type I IFN Alpha (PBL) was used at 5–500 U/mL. Lipofectamine 3000 (Thermo Fisher Scientific) was used to deliver cGAMP (InvivoGen). Z-AAD-CMK (Enzo Life Sciences) was dissolved in water at 10 mM and diluted in medium at a final concentration of 100 μM. Ruxolitinib (LC Laboratories) was dissolved in DMSO at 5 mM and then diluted in medium at a final concentration of 2.5 μM.
Chen J., Cao Y., Markelc B., Kaeppler J., Vermeer J.A, & Muschel R.J. (2019). Type I IFN protects cancer cells from CD8+ T cell–mediated cytotoxicity after radiation. The Journal of Clinical Investigation, 129(10), 4224-4238.
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2

IFN Alpha Treatment of Cells

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Cells were seeded into 6-well plates at 3×105 cells per well. The following day, cells were treated with 2 mL of DMEM/F-12 media supplemented with 10% FBS and 50 units/mL of Universal Type I IFN Alpha (PBL Assay Science Cat. #11200). The cells were harvested by aspirating the media, washing twice with 2 mL of sterile 1X PBS, then lysing with 350 μL RLT buffer from the RNeasy Mini kit (Qiagen). The cell lysate was stored at −80°C until RNA isolation was completed using the RNeasy kit following the manufacturer’s protocol.
De La Cruz-Rivera P.C., Kanchwala M., Liang H., Kumar A., Wang L.F., Xing C, & Schoggins J.W. (2017). The IFN response in bats displays distinctive ISG expression kinetics with atypical RNASEL induction. Journal of immunology (Baltimore, Md. : 1950), 200(1), 209-217.
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3

Viral Infection Quantification in HeLaS3 Cells

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Approximately 2.5 × 105 monolayer HeLaS3 cells were pre-treated with 500 IU/ml of IFN α(Universal type I IFN alpha, PBL Assay Science) for 24 h, washed twice with PBS, then infected by virus at m.o.i=0.01 for 40 min. Virus was removed and cells were washed with PBS and supplemented with fresh medium and incubated at 37 °C for 24 h. Plates were then frozen at −80 °C. Following 3 freeze and thaw cycles, plaque assays were performed on HelaS3 cells to quantify virus production.
Xiao Y., Dolan P.T., Goldstein E.F., Li M., Farkov M., Brodsky L, & Andino R. (2017). Poliovirus intrahost evolution is required to overcome tissue-specific innate immune responses. Nature Communications, 8, 375.
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4

IFN-β Promoter Luciferase Assay

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1.25 × 105 HEK293 cells/well were plated in 24‐well plates, transfected with 125 ng of p125‐Luc, encoding a Firefly luciferase reporter gene under the control of the IFN‐β promotor (gift from T. Fujita, Kyoto University, Japan) and 50 ng pRL‐TK (Promega) using Lipofectamine 2000, and treated with 200 U/ml universal type I IFN alpha (PBL Assay Science). Six‐eight hours later, cells were transfected with water (mock) or with the indicated amounts of total RNA extracted from HeLa cells that were infected with EMCV overnight (Fig 3A) or with RNA extracted from LGP2 immunoprecipitates (Fig 3B). Luciferase activity was measured 16–17 h later using the dual‐luciferase assay reporter system (Promega). Firefly luciferase values were divided by Renilla luciferase values to normalise for transfection efficiency. All data are shown as fold increase relative to mock‐transfected cells.
van der Veen A.G., Maillard P.V., Schmidt J.M., Lee S.A., Deddouche‐Grass S., Borg A., Kjær S., Snijders A.P, & Reis e Sousa C. (2018). The RIG‐I‐like receptor LGP2 inhibits Dicer‐dependent processing of long double‐stranded RNA and blocks RNA interference in mammalian cells. The EMBO Journal, 37(4), e97479.
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5

IFN-Induced Siglec-1 and Tetherin Expression

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Both M-CSF and GM-CSF matured MDMs were assayed for Siglec-1 and tetherin cell surface concentrations in the presence or absence of 1000 U/ml Universal Type I IFN Alpha (PBL Assay Science). Mature MDMs were stimulated overnight with IFN, cells washed with PBS and detached using Versene (Life Technologies) with gentle scraping. Surface CD14 (BD Pharmingen, Cat. No. 555399), sheep anti-Siglec-1 (R&D Systems, Cat. No. AF5197) and tetherin staining procedures were performed as previously described [35 (link)]. FACS Canto II flow cytometer (BD Biosciences) and FlowJo software (Treestar Inc) were used for analyses.
Hammonds J.E., Beeman N., Ding L., Takushi S., Francis A.C., Wang J.J., Melikyan G.B, & Spearman P. (2017). Siglec-1 initiates formation of the virus-containing compartment and enhances macrophage-to-T cell transmission of HIV-1. PLoS Pathogens, 13(1), e1006181.
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