Ifn α

Manufactured by PBL Assay Science

Sourced in United States

IFN-α is a type I interferon that plays a crucial role in the body's immune response. It is a cytokine produced by various cell types, including leukocytes, fibroblasts, and epithelial cells, in response to viral infections or other stimuli. IFN-α has antiviral, antiproliferative, and immunomodulatory properties, making it an important tool for researchers studying immune system function and disease processes.

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Lab products found in correlation

Ifn β, by PBL Assay Science (7 mentions) Interleukin 2 (il 2), by R&D Systems (4 mentions) Interleukin 6 (il 6), by R&D Systems (4 mentions) Ifn γ, by Thermo Fisher Scientific (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Tnf α, by R&D Systems (3 mentions) Ifn β, by BioLegend (3 mentions) Interleukin 6 (il 6), by BioLegend (3 mentions) Tnf α, by Thermo Fisher Scientific (3 mentions) Lipopolysaccharides (lps), by InvivoGen (3 mentions) Interleukin 4 (il 4), by R&D Systems (3 mentions) Ifn β, by R&D Systems (3 mentions) Il 17a, by BioLegend (3 mentions) Ifn γ, by BioLegend (3 mentions) Il 12, by Thermo Fisher Scientific (3 mentions) Il 12p40, by Thermo Fisher Scientific (3 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (3 mentions) Ip 10, by R&D Systems (2 mentions) Human xl cytokine array, by R&D Systems (2 mentions) Elisa kits to detect, by R&D Systems (2 mentions)

Spelling variants of this product

Mouse IFN-α (6 citations) VeriKine Human IFN-α Multi-Subtype ELISA Kit (4 citations) VeriKine Human IFN-α Multi-Subtype Serum ELISA kit (3 citations) Mouse IFN-α ELISA kit (3 citations) Universal type I IFN-α (2 citations) Anti-IFN-α (2 citations) FITC-conjugated anti-mouse IFN-α (2 citations) VeriKine‐HS Human IFN‐α All Subtype TCM ELISA Kit (2 citations) Human IFN-α (Alpha 2a) (2 citations) IFN-α and IFN-β (2 citations)

86 protocols using ifn α

NK Cell Degranulation Assay

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Fresh lung cells and matched PBMCs were resuspended in R10 medium and rested for 15 to 18 hours at 378C. For assays in which NK cells were primed with IFN-a, cryopreserved lung cells and PBMCs were thawed and incubated with 10 4 U/mL IFN-a (PBL Assay Science) for 15 hours before the degranulation assay. Subsequently, the cells were cocultured with R10 medium alone, K562 cells, or 721.221 cells and 1 mg/mL anti-CD20 mAb (Rituximab, Roche) for 2 hours in the presence of fluorescein isothiocyanate (FITC)-conjugated anti-CD107a (clone H4A3; BD Biosciences, San Jose, Calif).

Marquardt N., Kekäläinen E., Chen P., Kvedaraite E., Wilson J.N., Ivarsson M.A., Mjösberg J., Berglin L., Säfholm J., Manson M.L., Adner M., Al-Ameri M., Bergman P., Orre A.C., Svensson M., Dahlén B., Dahlén S.E., Ljunggren H.G, & Michaëlsson J. (2017). Human lung natural killer cells are predominantly comprised of highly differentiated hypofunctional CD69^-CD56^dim cells. The Journal of allergy and clinical immunology, 139(4).

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Interferon-α Sensitivity Assay

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Amounts of 3 × 10³ HT1080 cells were plated in a 96-well plate, transfected with siRNA SMARTpools or the most efficient individual siRNA (Dharmacon) (Table 1), treated with increasing concentrations of IFN-α or a fixed dose of 10 U/ml of IFN-α (catalog number 11200-2; PBL Assay Science) at 8 h.p.t., and harvested as described above.

Kueck T., Bloyet L.M., Cassella E., Zang T., Schmidt F., Brusic V., Tekes G., Pornillos O., Whelan S.P, & Bieniasz P.D. (2019). Vesicular Stomatitis Virus Transcription Is Inhibited by TRIM69 in the Interferon-Induced Antiviral State. Journal of Virology, 93(24), e01372-19.

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Quantifying Cytokine Levels in Samples

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CXCL10 ELISA was performed using commercially available paired antibodies and recombinant cytokines (Becton Dickenson, Franklin Lakes, NJ); the limit of detection was 15.6 pg/ml. IFN-α (PBL Interferon Source, Piscataway, NJ) was assayed via commercial ELISA kit according to the manufacturer’s instructions; the IFN-α “multi-subtype” kit detects all isoforms except IFNα₁₃ and IFNα₂₁. The limit of detection was 9.7 pg/ml. Active Motif TransAM Kits and the Cayman Chemical NF-κB (human p50/p65) Combo Transcription Factor Assay Kits were used as per the manufacturers’ protocols.

Pritchard A.L., White O.J., Burel J.G., Carroll M.L., Phipps S, & Upham J.W. (2014). Asthma Is Associated with Multiple Alterations in Anti-Viral Innate Signalling Pathways. PLoS ONE, 9(9), e106501.

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Quantifying Cytokine Secretion in NPC

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Cell culture supernatants from NPC were collected 24 h after stimulation with TLR1–9 ligands and tested for secretion of IL6, TNF and IL10 by enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions (R&D systems). Cell culture supernatants were further analyzed for secretion of IFNA (PBL Assay Science), IFNB1 (PBL Assay Science), IFNL1 (eBioscience) and IFNL2/3 (RayBiotech, Norcross, GA, USA).

Werner M., Schefczyk S., Trippler M., Treckmann J.W., Baba H.A., Gerken G., Schlaak J.F, & Broering R. (2022). Antiviral Toll-like Receptor Signaling in Non-Parenchymal Liver Cells Is Restricted to TLR3. Viruses, 14(2), 218.

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Cell Line Maintenance and Testing

Cited 1 time

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HT1080 cells were obtained from ATCC and maintained in Dulbecco’s modified Eagle’s medium (DMEM) (catalog number 11995065; Thermo Fisher) with 10% fetal calf serum (FCS) (catalog number F8067-500ML; Sigma) and gentamicin (catalog number 15710064; Thermo Fisher). An IFN-α (catalog number 11200-2; PBL Assay Science)-sensitive HT1080 single-cell clone was derived by limiting dilution. BHK21 cells obtained from ATCC were maintained in Eagle’s minimum essential medium (EMEM) (catalog number 30-2003; ATCC) supplemented with 10% FCS (catalog number F8067-500ML; Sigma) and gentamicin (catalog number 15710064; Thermo Fisher). Cells were not tested for mycoplasma contamination but were periodically tested for retrovirus contamination using a PCR-based reverse transcriptase assay (35 (link), 36 (link)).
BSRT7 cells (a kind gift from K. Conzelmann) (37 (link)) and African green monkey kidney Vero cells (CCL-81; ATCC) were maintained in DMEM (catalog number 10-013-CV; Corning, Inc.) containing 10% fetal bovine serum (FBS) (catalog number TCB 101; Tissue Culture Biologicals) at 37°C and 5% CO₂.

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Quantification of Inflammation Biomarkers in CVL

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Inflammation biomarkers were quantified in CVL from Group 2 women using a Milliplex 17-plex kit (Millipore) run on Luminex
MagPix (GM-CSF, IFN-γ, IL-10, IL-12p40, IL-12p70, IL-15, IL-1RA, IL-1α, IL-2, IL-4, IL-6, IL-8, IP10, MCP-1,
MIP-1β, RANTES, and TNFα). Five additional biomarkers were tested by ELISA: CD163 (R&D Systems), SLPI (R&D
Systems), IFN-α (PBL Assay Science), and beta-defensins 2 and 3 (Assay Biotech).

Nelson J.A., PARIS K.D., RAMIREZ C., EDMONDS A., MOLLAN K.R., BAY C.P., COMPLIMENT K., HEROLD B.C., ANASTOS K., MINKOFF H., KASSAYE S., SEIDMAN D.L., FRENCH A.L., GOLUB E.T., SHETH A.N., OCHSENBAUER C., SWANSTROM R., ERON J.J, & ADIMORA A.A. (2020). Female Genital Tract Shedding of HIV-1 is Rare in Women with Suppressed HIV-1 in Plasma. AIDS (London, England), 34(1), 39-46.

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Evaluation of Plasma Immune Markers

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Presence of plasma IFNα (PBL Assay Science), IP-10 (R&D systems), sCD14 (R&D systems), TNFα (R&D systems), D-Dimer (Abcam), and C-Reactive Protein (CRP) (Abcam) was evaluated by ELISA per manufacturers protocol.

Saxena M., Sabado R.L., La Mar M., Mohri H., Salazar A.M., Dong H., Correa Da Rosa J., Markowitz M., Bhardwaj N, & Miller E. (2019). Poly-ICLC, a TLR3 Agonist, Induces Transient Innate Immune Responses in Patients With Treated HIV-Infection: A Randomized Double-Blinded Placebo Controlled Trial. Frontiers in Immunology, 10, 725.

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Cytokine Profiling of IFN-β Deficient BMDMs

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IFN-β^−/− BMDMs were infected as described previously and treated with 50 pg/mL IFN-β. An additional 50 pg/mL of IFN-β was added at 3 hpi, and cell culture supernatants were collected at 6 hpi. Concentrations of selected cytokines were determined using a custom ProcartaPlex magnetic bead-based multiplex assay (Thermo Fisher Scientific) on the Luminex MAGPIX^® platform according to manufacturer’s instructions. IFN-α (PBL Assay Science), IFN-β (BioLegend), CCL12 (R&D Systems) and CCL3 (R&D Systems) protein levels were measured using ELISAs.

Banks D.A., Ahlbrand S.E., Hughitt V.K., Shah S., Mayer-Barber K.D., Vogel S.N., El-Sayed N.M, & Briken V. (2019). Mycobacterium tuberculosis inhibits autocrine type I interferon signaling to increase intracellular survival.. Journal of immunology (Baltimore, Md. : 1950), 202(8), 2348-2359.

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Vaccination-Induced Immune Responses

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Serum from whole blood, spleens and popliteal lymph nodes were collected at specified time points after vaccination. Supernatants were collected from single cell suspensions of spleens and LNs that were cultured in vitro for 12 hours in complete RPMI at 37°C. Commercially available ELISA kits were used to measure IL-12p40 (Peprotech) and all subtypes of IFN-α (PBL Assay Science) as per manufacturer’s protocols.

Baharom F., Ramirez-Valdez R.A., Tobin K.K., Yamane H., Dutertre C.A., Khalilnezhad A., Reynoso G.V., Coble V.L., Lynn G.M., Mulè M.P., Martins A.J., Finnigan J.P., Zhang X.M., Hamerman J.A., Bhardwaj N., Tsang J.S., Hickman H.D., Ginhoux F., Ishizuka A.S, & Seder R.A. (2020). Intravenous Nanoparticle Vaccination Generates Stem-Like TCF1+ Neoantigen-Specific CD8+ T Cells. Nature immunology, 22(1), 41-52.

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Cytokine Profiling of Viral Infection

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Cell supernatants were collected 24 or 72 hr after infection and frozen at −80°C. Thawed samples were applied to Human XL Cytokine Array (ARY022, R&D Systems) per kit protocols. For increased sensitivity, the maximum volume of supernatant (500 µl) was used on the array. ELISA kits to detect TNFα, CCL2, and CXCL3 (R&D Systems) or IFNα (PBL Assay Science) were used according to the manufacturer’s protocol.

Hanners N.W., Eitson J.L., Usui N., Richardson R.B., Wexler E.M., Konopka G, & Schoggins J.W. (2016). Western Zika Virus in Human Fetal Neural Progenitors Persists Long Term with Partial Cytopathic and Limited Immunogenic Effects. Cell reports, 15(11), 2315-2322.

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