Human Lipocalin-2/NGAL Quantikine ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all reagents to room temperature before use. Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution. -Working dilutions should be prepared and used immediately.	-A standard curve should be generated for each set of samples assayed. -It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum. -Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.

Upstream tips
-Bring all reagents to room temperature before use. Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution. -Working dilutions should be prepared and used immediately.
Protocol tips
-A standard curve should be generated for each set of samples assayed. -It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum. -Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.

Publication protocol

Firstly, four milliliter venous blood was withdrawn from SZ patients and the corresponding controls in the morning into procoagulant tube prior to administration of any medication. Serum was separated by centrifugation (3500 r/min, 5 min) from coagulated blood, then was collected and stored at -80℃ until analysis. Serum NRG1, ErbB4, BDNF, DNMT1 and TET1 protein levels were measured by commercially available ELISA kits (NRG1β1/ErbB4/BDNF, R&D Systems, Minneapolis MN; DNMT1/TET1, Cusabio, Wuhan, China) following the manufacturer's instructions. The 96-well micro plates were incubated overnight with monoclonal antibody at 4℃. Samples and standard proteins were added after incubation with blocking sample buffer. Plates were then treated with enzyme-labeled polyclonal antibody. Then, H2O2 was added and the color was developed after addition of TMB solution. After adding 2 mol/L H2SO4 to stop the reaction, the absorbance at 450 nm were measured on micro plate reader. Protein concentrations were determined according to the standard curve.

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Manufacturer protocol

Download the product protocol from R&D Systems for Human Lipocalin-2/NGAL Quantikine ELISA Kit below.

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