High glucose Dulbecco’s modified Eagle’s Medium (DMEM, GIBCO, PA, USA). Fetal bovine serum (FBS, EVERY GREEN, China). Penicillin/streptomycin (P/S, GIBCO, USA). AG490 (HY-12000, MCE). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) (Solarbio, Beijing, China). Docetaxel (114977–28-5, aladdin, shanghai, china). IL-6 (Mouse IL-6 Flex Set, 558301, BD), IL-17A (Mouse IL-17A Flex Set, 560283, BD), TNF-α (Mouse TNF Flex Set, 558299, BD), IFN-γ (Mouse IFN-γ Flex Set, 558296, BD). DAPI staining reagent (G1012, servicebio, Wuhan, China). SZYY obtained from C. officinalis Planting Base in Taiping Town, Xixia County, Nanyang City, Henan Province, China.
Mouse tnf flex set
Manufactured by BD
The Mouse TNF Flex Set is a laboratory equipment product designed to measure tumor necrosis factor (TNF) levels in mouse samples. It provides a standardized and reliable method for quantifying TNF in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Mouse il 6 flex set, by BD (3 mentions)
Mouse il 10 flex set, by BD (3 mentions)
Mouse ifn γ flex set, by BD (2 mentions)
Mouse il 17a flex set, by BD (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Cytometric bead array cba, by BD (1 mentions)
Flat 96 well plates, by Corning (1 mentions)
Mouse mcp 1 flex set, by BD (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
Human tnf flex set, by BD (1 mentions)
Cytometric bead array, by BD (1 mentions)
Human il 10 flex set, by BD (1 mentions)
Human il 6 flex set, by BD (1 mentions)
5 protocols using mouse tnf flex set
1
Anti-tumor Effects of SZYY in Cells
2
Cytokine Quantification in Cell Supernatant
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To assess cell supernatant cytokine concentrations, the spleen cell suspensions were centrifuged and supernatants collected after a 24-h incubation time. The concentrations of IL-6 and TNF-α were measured using the BD Cytometric Bead Array Mouse IL-6 Flex Set and Mouse TNF Flex Set (BD Biosciences) according to the manufacturer’s instructions.
3
Quantification of Nitric Oxide and Cytokines
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The measurement of nitric oxide (NO) in the supernatants was taken using the modified Griess reagent (Sigma, G4410). Samples (100 μl) were plated in a flat 96‐well plates (Corning), mixed with equal volumes of freshly prepared Griess reagent, and incubated for 15 min at room temperature. The plate was read at 540 nm using a microplate reader (LabSystems). The cytokine release to the supernatants was measured using BD™ Cytometric Bead Array (CBA) following the manufacturer’s protocol and detected using a BD FACSVerse flow cytometer. The following kits were used to detect indicated cytokines: Mouse IL‐6 Flex Set (BD, 558301), Mouse TNF Flex Set (BD, 558299), Mouse MCP‐1 Flex Set (BD, 558342), and Mouse IL‐10 Flex Set (BD, 558300). The cytokine level in the supernatant was reflected by the median fluorescence intensity (MFI) of PE of the corresponding capture beads.
4
Cytokine Quantification in Mouse Serum
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The contents of IL-10 (Mouse IL-10 Flex Set, 558300, BD), IFN-γ (Mouse IFN-γ Flex Set, 558296, BD), TNF (Mouse TNF Flex Set, 558299, BD), and IL-2 (Mouse IL-2 Flex Set, 558297, BD) in the serum of mice were detected using CBA according to the manufacturer’s protocols. Initially, the different concentrations of standard products were formulated according to the respective preparations, and a standard curve was plotted. Then, 50 μL of serum and 50 μL of microspheres were mixed and incubated at room temperature in the dark for 2 h. Then, 50 μL of PE antibody conjugated with TNF, IL-2, IL-1β, and IFN-γ was added and incubated for 1 h at room temperature in the dark. Further, 1 mL of washing solution was added and centrifuged, and the supernatant was gently discarded. The resulting suspension was resuspended in 400 μL of the buffer, and finally, the cytokine levels were measured by FCM, and the results were analyzed using the Diva (BD).
5
Cytokine profiling in humanized mice
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Plasma samples of humanized mice were stored at -80 C. After defrosting, levels of human and mouse TNF-a, IL-6 and IL-10 were analyzed using Cytometric Bead Array (all from BD Biosciences: CBA Mouse TNF Flex Set, 558299; CBA Mouse IL-6 Flex Set, 558301; CBA Mouse IL-10 Flex Set, 558300; CBA Human TNF Flex Set, 558273; CBA Human IL-6 Flex Set, 558276; CBA Human IL-10 Flex Set, 558274) according to the manufacturer's instructions. The lower detection limits were: 1.2 pg/ ml, human TNF-a; 2.8 pg/ml, murine TNF-a; 1.6 pg/ ml, human IL-6; 1.4 pg/ml, murine IL-6; 0.13 pg/ml, human IL-10; 9.6 pg/ml, murine IL-10. If measured values were below these detection limits, they were set to zero.
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