Human PAI-1 PicoKine™ ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store at 4°C for 6 months, at -20°C for 12 months. -Avoid multiple freeze-thaw cycles. -The standard solutions are best used within 2 hours.	-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment. -Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature. -Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection.

Upstream tips
-Store at 4°C for 6 months, at -20°C for 12 months. -Avoid multiple freeze-thaw cycles. -The standard solutions are best used within 2 hours.
Protocol tips
-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment. -Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature. -Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection.

Publication protocol

Patients were instructed in 24-h timed urine collection for the measurement of UAER. Urinary albumin was determined by calorimetric methods using commercial kits (ZiestChem Diagnostics, Tehran, Iran). Blood samples were collected after 12 h of overnight fasting, and biochemical analysis was made subsequently. Glucose measurements were made using glucose oxidase method (intra-assay coefficient of variants (CV) = 2.1 %, inter-assay CV = 2.6 %). Creatinine was measured using calibrated Jaffe method (Parsazmoon, Karaj, Iran). Total cholesterol, HDL-C, low-density lipoprotein cholesterol (LDL-C), and triglycerides were determined using direct enzymatic methods (Parsazmoon, Karaj, Iran). HbA1c was estimated by high-pressure liquid chromatography. HSPA1A was measured using a quantitative sandwich ELISA immunoassay (EKS-715, Stressgen). The intra- and inter-assay CVs were 4.5 and 7 %, respectively. Apo-A1 was measured (Cobas INTEGRA Tina-quant Apolipoprotein A-1 ver.2) following the principles of antigen-antibody reaction using immunoturbidometric method (intra-assay CV = 0.8 %, inter-assay CV = 1.7 %). PAI-1 was measured by sandwich ELISA (Human PAI-1 PicoKine ELISA kit, EK0859, Boster Bio, CA, USA; intra-assay CV = 5–6.5 %, inter-assay CV = 6.4–8.2 %).

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Manufacturer protocol

Download the product protocol from BosterBio for Human PAI-1 PicoKine™ ELISA Kit below.

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