Human il 8 quantikine elisa kit

Circulating levels of IL-8 were measured in sera from all RTHα patients and controls using the Human IL-8 Quantikine ELISA kit (R&D Systems) according to manufacturer’s instructions with the following modifications: sample volume was 100 µL, incubation time was 3 h and the following points were added to the standard curve: 3.6, 7.7, 15.8 and 31.3 pg/mL. Samples were measured in duplicate and samples below the detection limit (7.7 pg/mL) were assigned a value of half the detection limit.
A panel of pro-inflammatory cytokines and chemokines (IL-1β, IL-6, TNF and IL-8) were measured in supernatant of stimulated macrophages and in plasma from the adult RTHαD211G patient and healthy controls using the Human Inflammatory Cytokine Cytometric Bead Array kit (BD Biosciences, Franklin Lakes, NJ, USA). Samples were run in triplicate on a FACS Calibur flow cytometer (BD Biosciences). All samples were measured in the same run. Data were analyzed using FlowJo software (version 10).

Serum levels of human IL-8 were detected by using Human IL-8 Quantikine ELISA Kit (R&D Systems, Inc.) follows the instructions.

The level of IL-8 was quantified using human IL-8 Quantikine ELISA Kit from R&D Systems (Minneapolis, MN), according to the manufacturer’s instructions.

Plasma BDNF levels were measured using a BDNF Kit (Quantikine Human BDNF kit; R&D Systems, Minneapolis, MN, USA) (www.RnDSystems.com) and an ELISA reader with a minimum detectable dose of 80 pg/mL (SpectraMax-M2; Molecular Devices, Sunnyvale, CA, USA). All samples were analyzed twice. The cytokine ELISA kits used were also purchased from R&D systems and included a Human CRP Quantikine ELISA Kit, a Human TNF-alpha Quantikine ELISA Kit, and a Human IL-8 Quantikine ELISA Kit for the assessment of plasma levels of CRP, TNF-α, and IL-8, respectively.

After the preparation of the young and senescent cell model as described above and treatment with ADC and controls, the culture media were collected and subjected to ELISA [Human IL-6 Quantikine ELISA Kit (D6050), Human IL-8 Quantikine ELISA Kit (D 8000C), Human MMP-9 Quantikine ELISA Kit (DMP900) (R & D Systems, Inc., Minneapolis, MS, USA), and Human MMP3 ELISA Kit (ab100607, abcam)] for quantifying IL-6, IL-8, MMP9, and MMP3 concentration.

The CM of HSC-2, SQUU-A, and SQUU-B cells were examined using the human IL-8 Quantikine ELISA Kit (R&D Systems) and human SerpinE1 Quantikine ELISA Kit (R&D Systems) in accordance with the manufacturer’s guidelines. RPMI1640 with 2% FBS served as the negative control. IL-8 and SerpinE1 concentrations were measured using the Multiskan FC microplate reader (Thermo Fisher Scientific, Waltham, MA, USA) at a wavelength of 450 nm.