Rat Superoxide dismutase [Mn], mitochondrial(SOD2) ELISA kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Bring all reagents to room temperature (18-25°C) before use for 30min. -Prepare fresh standard for each assay. Use within 4 hours and discard after use. -Centrifuge the sample again after thawing before the assay.	- To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. -Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. -TMB Substrate should remain colorless or light blue until added to the plate. Please protect it from light. -Stop Solution should be added to the plate in the same order as the TMB Substrate.

Upstream tips
Bring all reagents to room temperature (18-25°C) before use for 30min. -Prepare fresh standard for each assay. Use within 4 hours and discard after use. -Centrifuge the sample again after thawing before the assay.
Protocol tips
- To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. -Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. -TMB Substrate should remain colorless or light blue until added to the plate. Please protect it from light. -Stop Solution should be added to the plate in the same order as the TMB Substrate.

Publication protocol

Kidney tissues were placed in labeled glass tubes. The samples were washed with phosphate‐buffered saline (PBS) once and stored at −80°C until biochemical analysis. The samples were thawed before the analysis. All the tissues were homogenized once in 1 mL of PBS. The samples were frozen and thawed twice to facilitate cell membrane disintegration. The homogenate was centrifuged at 4°C at 5000g for 5 minutes, and the supernatant was removed. Catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH‐Px) activities were measured using commercially available enzyme‐linked immunosorbent assay kits (Cusabio Biotech, Wuhan, PRC), according to the manufacturer's instructions. Malondialdehyde (MDA) concentrations were measured using commercially available colorimetric assay kits (Cayman Chemical Company, State, U.S.A.), according to the manufacturer's instructions. The bicinchoninic acid (BCA) protein assay was used for quantitation of the total protein in the tissue (Thermo Fisher Scientific Inc., Rockford, State, U.S.A.).

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Manufacturer protocol

Download the product protocol from Cusabio for Rat Superoxide dismutase [Mn], mitochondrial(SOD2) ELISA kit below.

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