Assay Principle:

The DetectX hours and allows for flexible sample input based on the anticipated sensitivity of the assay.

Users can opt for 50 μL of input for an expected sensitivity of 20.9 pg/mL or 100 μL for 14.4 pg/mL. Detailed instructions are available in the kit insert, which should be read thoroughly before running the assay. The kit includes a corticosterone standard to create a precise standard curve.

Protocol Summary:

• Transfer standards or diluted samples into the provided transparent microtiter plate coated with donkey anti-sheep IgG antibody.
• Add corticosterone peroxidase conjugate and corticosterone polyclonal sheep antibody to initiate the immunological reaction.
• Incubate the mixture for 1 hour at room temperature with shaking. The reaction varies inversely with the corticosterone concentration in the sample.
• After incubation, wash away excess conjugate and add the TMB substrate. This substrate reacts with the bound conjugate, creating a measurable signal.
• Use a plate reader to detect the signal at 450nm and calculate corticosterone concentration using the standard curve.

Background:

Corticosterone, also known as Kendall;fight-or-flight” response and chronic elevations linked to various health issues. As a memory-modulating hormone capable of crossing the blood-brain barrier, corticosterone interacts with multiple brain regions, influencing memory consolidation processes. Elevated levels occur in response to severe burn injuries and drastic dietary restrictions. Beyond its role in stress response, corticosterone affects sleep-wake patterns, supporting alertness and wakefulness, and plays critical roles in the developmental and behavioral processes in birds.

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