The IRAP (Insulin-Regulated Aminopeptidase) ELISA Kit is designed to quantitatively measure the levels of Insulin-Regulated Aminopeptidase (IRAP) in biological samples. IRAP is a key enzyme involved in the processing of various peptides, including neuropeptides, and plays a significant role in glucose metabolism, insulin signaling, and other physiological processes.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to IRAP, allowing for capture of IRAP from the samples.
- Standards:
- Solutions containing known concentrations of recombinant or purified IRAP, used to generate a standard curve for quantifying IRAP levels in samples.
- Sample Diluent:
- A buffer solution used to dilute biological samples to appropriate concentrations for accurate measurement.
- Detection Antibody:
- An antibody specific to a different epitope of IRAP, often conjugated to an enzyme such as HRP for detection.
- Enzyme Conjugate:
- If the detection antibody is biotinylated, a streptavidin-HRP conjugate may be included for signal detection.
Procedure
- Blocking:
- Block any remaining uncoated surfaces in the wells with a blocking buffer to prevent non-specific binding.
- Addition of Standards and Samples:
- Add standards and samples to the wells. The IRAP in the samples will bind to the immobilized antibody.
- Incubation:
- Incubate the plate to allow IRAP to bind to the antibody.
- Addition of Detection Antibody:
- Add a detection antibody specific to IRAP, which binds to the captured IRAP.
- Washing:
- Wash the wells to remove unbound substances, including excess detection antibodies.
Performance Characteristics
- Sensitivity: Capable of detecting low levels of IRAP, typically in the picogram per milliliter (pg/mL) range.
- Precision: Shows low intra-assay and inter-assay variability, with coefficients of variation (CV) generally below 10%.
- Linearity: Demonstrates linearity across the standard curve range, ensuring accurate quantification.
- Recovery: High recovery rates, usually above 85%, indicating reliable performance across different sample types.
Applications
- Metabolic Research: Studying the role of IRAP in insulin signaling and glucose metabolism.
- Neuropeptide Processing: Investigating IRAP’s role in processing neuropeptides and its implications in neurological disorders.
- Diabetes Research: Assessing changes in IRAP levels related to diabetes and metabolic syndrome.
- Cancer Research: Exploring the involvement of IRAP in cancer progression and metastasis.
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