Rat Retinol binding protein 4,RBP-4 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

The serum insulin level (Mercodia Co., Sweden) and gastrocnemius RBP4 content (Cusabio Biotech Co., Ltd. Sino-American) were determined using commercial enzyme-linked immunosorbent assay kits. All analyses were performed in accordance with the manufacturers’ recommendations. The sensitivities of the kits were 0.015 μg/L and 15.6 ng/ml for insulin and RBP4, respectively. Homeostasis model assessment-estimated insulin resistance (HOMA-IR) was calculated according to the following equation: [glucose (mg/dl) × insulin (μU/ml)/405].

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Manufacturer protocol

Download the product protocol from Cusabio for Rat Retinol binding protein 4,RBP-4 ELISA Kit below.

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