ELISA kits for detection of IFN-α (42120-1), IFN-β (MIFNB0), IFN-γ (MIF00), TNF-α (MTA00B), IL-2 (M2000), IL-6 (M6000B), IL-10 (M1000B), IL-12 (M1270), and IL-23 (M2300) were purchased from R&D System. ELISA kits for detection of TGF-β (ab119557), IL-1α (ab199076), IL-1β (ab100705), and IL-4 (ab100710) were purchased from Abcam. Cytokine levels were determined using corresponding ELISA kits according to the manufacturer’s instructions.
Mifnb0
Manufactured by R&D Systems
MIFNB0 is a recombinant human Macrophage Migration Inhibitory Factor (MIF) protein. MIF is a pro-inflammatory cytokine involved in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Mif00, by R&D Systems (2 mentions)
Difnb0, by R&D Systems (2 mentions)
Mta00b, by R&D Systems (2 mentions)
Ab119557, by Abcam (1 mentions)
Ab199076, by Abcam (1 mentions)
Ab100705, by Abcam (1 mentions)
Ab100710, by Abcam (1 mentions)
M2300, by R&D Systems (1 mentions)
Mouse ifn beta quantikine, by R&D Systems (1 mentions)
Sm2000, by R&D Systems (1 mentions)
Mouse ifn β, by PBL Assay Science (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Universal master mix 2 with ung, by Thermo Fisher Scientific (1 mentions)
Hiscript 3 rt supermix, by Vazyme (1 mentions)
Gdna wiper, by Vazyme (1 mentions)
10 protocols using mifnb0
1
Cytokine Detection Using ELISA
2
Quantifying Cytokine Levels in Mice
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Interferon beta (IFNB; R&D Systems, MIFNB0 [mouse] or DIFNB0 [human]) or tumor necrosis factor-alpha (TNFα; R&D Systems, MTA00B [mouse]) in the plasma of mice obtained from retro-orbital blood and in the culture supernatants of mouse or human aortic EC were measured according to the manufacturer’s protocol using an ELISA.
3
IFNβ Secretion Quantification in Keratinocytes
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IFNβ secretion from Cre and CKOβ primary keratinocytes was measured in cell supernatants using Mouse IFN beta Quantikine enzyme-linked immunosorbent assay (ELISA) kit (MIFNB0, R&D Systems, Inc.). IFNβ concentrations were derived from the absorbance values measured at 450 nm with a correction at 540 nm using the Multiskan EX microplate spectrophotometer (ThermoFisher) and converted to concentration values based on the standards provided from each kit.
4
IFN-β Production in RSV-Stimulated Alveolar Macrophages
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ALMs (1 × 106/mL) were stimulated with RSV (MOI 1) or mock-stimulated for 1, 4 or 24 h at 37 °C under 5% CO2. Alternatively, ALMs were stimulated with TLR3 ligand Poly I:C (10 and 20 μg/mL) for 24 h at 37 °C under 5% CO2. Afterwards, supernatants were collected, and IFN-β concentrations were determined using ELISA (R&D Systems, cat MIFNB0), following the manufacturer’s instructions.
5
Cytokine Profiling in Cell Culture
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6
Measurement of Cytokine Secretion in MEFs
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MEFs were transfected with poly(I:C)-LMW (1 μg/mL), poly(I:C)-HMW (1 μg/mL), or poly(dI:dC) (1 μg/mL), or stimulated with LPS (100 ng/mL) or bovine serum albumin (BSA)-conjugated palmitate (800 μM) for 8 h. The culture supernatants were collected and centrifuged to remove nonadherent cells. The concentrations of mouse IFN-β (#42400-1; PBL Assay Science, Piscataway, NJ, or MIFNB0; R&D Systems, Minneapolis, MN) and mouse IL-6 (M6000B; R&D Systems) were determined by ELISA according to the manufacturers’ instructions. The supernatants of SeV-infected MEFs were also examined.
7
Quantification of CXCL10 and IFN-β in Tumor Tissue
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The chromogenic reaction involving biotin-labeled and peroxidase-labeled avidin, catalyzed by peroxidase, was employed. CXCL10 (DY466-05, R&D Systems) and IFN-β (MIFNB0, R&D Systems) were separately detected using a sandwich enzyme-linked immunosorbent assay kit. Tumour tissue samples were sectioned into small pieces, and a lysis buffer containing protease inhibitors was added. The samples were homogenized with a homogenizer. After centrifugation, the supernatant was collected and analysed in accordance with the manufacturer’s instructions.
8
Cytokine and Chemokine Detection in BMDC and BMDM
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The supernatants of BMDC and BMDM were harvested for cytokine and chemokine detection. Murine IFN-β level was measured by enzyme-linked immunosorbent assay (ELISA) (MIFNB0, R&D). Other murine cytokines and chemokines were detected by multiplex fluorescence-encoded beads (741,044, BioLegend; 740,451, BioLegend).
9
Intratumoral Cytokine Profiling in EMT-6 Mice
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BALB/c mice were challenged with 1 × 105 EMT-6 cells. Tumor-bearing mice were treated with a single dose of 50 mg/kg MSA-2 when tumor volume reached 300 mm3. Six hours after MSA-2 treatment, mice were killed and tumors were collected for intratumoral cytokine and chemokine detection. The gross protein level of tumor tissue homogenate was measured by BCA protein assay kit (P0010S, Beyotime). The concentrations of cytokines and chemokines in the homogenate were detected by ELISA (MIFNB0, R&D) and multiplex fluorescence-encoded beads (741,044, BioLegend; 740,451, BioLegend).
10
RNA Extraction and RT-qPCR Analysis of Tumor Cells
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The total RNA of tumor cells and tumor tissues was extracted using an RNA Quick Purification Kit (Yishan Biotech) according to the manufacturer’s instructions. The RNA was then reverse-transcribed to complementary DNA (cDNA) using HiScript III RT SuperMix for real time quantitative PCR (RT-qPCR) with gDNA wiper (Vazyme, R323-01). SYBR Green III (Vazyme, Q711-02/03) was used for the RT-qPCR of non-TaqMan primers, and Universal Master Mix II with UNG (Thermo Fisher, 4440038) was used for the RT-PCR of TaqMan primers. The primer sequences are listed in online supplemental table 1 .
Supernatant levels of human IFN-β (R&D Systems, DIFNB0) and mouse IFN-β (R&D Systems, MIFNB0) were measured by ELISA kits following the manufacturer’s instructions.
Supernatant levels of human IFN-β (R&D Systems, DIFNB0) and mouse IFN-β (R&D Systems, MIFNB0) were measured by ELISA kits following the manufacturer’s instructions.
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