ELISA Rat - RBP4

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

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Rat Retinol binding protein 4,RBP-4 ELISA Kit

Cusabio

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.

Manufacturer protocol Publication protocol 1 Relevant paper

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