Human il 17a high sensitivity elisa kit

All patients were treated initially with a topical application of 5% salicylic acid ointment for desquamation. Plaque scales were collected from the most representative lesions. The same amount of scale from each patient was weighed. Then, plaque scales were suspended in phosphate-buffered saline (PBS) without calcium and magnesium and disrupted with Sonics Vibracell VCX130 in impulsing mode. Cell suspension was centrifuged for 10 minutes at 5000 x g. Then, scale samples were subdivided into small aliquots to be stored at -80°C until tested for cytokine levels. The concentration of IL-17 in the patient plaque scales was determined with the use of Human IL-17A High Sensitivity ELISA kit (eBioscience, Vienna, Austria) and Human IL-17 ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturers' instructions. With Human IL-17A High Sensitivity ELISA kit, we have reached very high results, which were out of range. That is why we repeated the analysis with Human IL-17 ELISA kit (R&D Systems, Minneapolis, MN, USA).

Blood samples were collected from psoriatic patients and controls and were centrifuged for 15 minutes at 1000 x g. Then, serum samples were subdivided into small aliquots to be stored at -80°C until tested for cytokine levels. In the studied psoriatic patients as well as the control group, concentrations of IL-17 were determined with the use of Human IL-17A High Sensitivity ELISA kit (eBioscience, Vienna, Austria), according to the manufacturer's instructions.

A 10-mL sample of peripheral venous blood was drawn from patients and healthy subjects into tubes that contained lithium heparin. Blood was incubated with the mAbs M5E2 against the molecule CD14 conjugated with peridinin chlorophyll protein (PerCP) and 3G8 against the molecule CD16 conjugated with FITC. Both, antibodies and the appropriate isotype controls were provided by Becton Dickinson (San Jose, CA). Flow cytometric analysis was performed with a FACScalibur flow cytometer (Becton Dickinson). Absolute CD14⁺CD16⁺⁺ monocyte numbers were obtained using BD TruCount Tubes (Becton Dickinson). To calculate the median fluorescence intensity (MFI) of the receptors, the flow cytometer was calibrated with BD Calibrite 3 beads (Becton Dickinson) to adjust set fluorescence compensation. Serum levels of interleukin-17 (IL-17) and Type II interferon y (IFNy) were analyzed by high sensitivity ELISA using the Human IL-17A High Sensitivity ELISA kit (eBioscience, Bender MedSystem, Vienna, Austria) and Human IFN-y High Sensitivity ELISA kit (eBioscience) in a subgroup of patients. The samples were measured using a Power Wave XS microplate reader (Biotek, VT) set to 450 nm. Patients were grouped according to whether their plasma levels of IL-17A were higher or lower the reference values for the healthy population (reference value <0.02 pg/ml).