Human Lipocalin-2/NGAL 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

The ELISA method was used for this. Wuhan Boster Biological Technology. Ltd. Kits (China) and associated standards were also used. At the time of measurement, urine samples stored at −80 °C were thawed at room temperature. A microplate coated with purified NGAL antibodies was used and a solid carrier was prepared. NGAL was successively added to monoclonal antibody-coated microplates, which were bound to HRP-labelled NGAL antibodies, forming antibody-antigen-enzyme-labelled-antibody complexes. TMB colorimetric substrate was added after thorough washing. For this assay, TMB turns blue with the catalytic action of HRP, and is converted to a yellow colour after the addition of acid. The depth of colour in the samples is directly proportional to the amount of NGAL in the sample. The absorbance at 450 nm (OD value) was measured using a spectrophotometer, and the NGAL concentration of the sample was then calculated.

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Manufacturer protocol

Download the product protocol from BosterBio for Human Lipocalin-2/NGAL PicoKine™ ELISA Kit below.

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