Rat Angiopoietin 2,ANG-2 ELISA Kit

Protocol tips

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	ELISA kits (R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer's instructions. Samples in 96-well microplates were read by a Benchmark plus microplate spectrophotometer (Bio-Rad Laboratories, Inc.) within 20 min after the reaction was stopped. Test samples and standards were measured in duplicate. Microplate manager version 5.2.1 software (Bio-Rad Laboratories, Inc.), which was equipped with the spectrophotometer, automatically calculated sample concentrations according to corresponding standard curves with four-parameter logistics and log-logit function.

Protocol tips

ELISA kits (R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer's instructions. Samples in 96-well microplates were read by a Benchmark plus microplate spectrophotometer (Bio-Rad Laboratories, Inc.) within 20 min after the reaction was stopped. Test samples and standards were measured in duplicate. Microplate manager version 5.2.1 software (Bio-Rad Laboratories, Inc.), which was equipped with the spectrophotometer, automatically calculated sample concentrations according to corresponding standard curves with four-parameter logistics and log-logit function.

Publication protocol

The kidney tissues' total protein content was measured by a Coomassie Brilliant Blue Total Protein Quantification kit following homogeneity (5 min, 0°C) and centrifugation (3,000 × g, 10 min, 4°C). Kidney proteins ACE, ACE2, Ang (1–7), Ang II, AT 1 and AT 2 were measured by rat ELISA kits (R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer's instructions. Samples in 96-well microplates were read by a Benchmark plus microplate spectrophotometer (Bio-Rad Laboratories, Inc.) within 20 min after the reaction was stopped. Test samples and standards were measured in duplicate. Microplate manager version 5.2.1 software (Bio-Rad Laboratories, Inc.), which was equipped with the spectrophotometer, automatically calculated sample concentrations according to corresponding standard curves with four-parameter logistics and log-logit function.

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