The ADCY (Adenylate Cyclase) ELISA Kit is designed for the quantitative measurement of adenylate cyclase (ADCY) levels in various biological samples, such as tissue homogenates, cell lysates, and possibly serum or plasma. Adenylate cyclase is an enzyme that converts ATP to cyclic AMP (cAMP) in response to activation by G-protein-coupled receptors (GPCRs). This cAMP signaling pathway plays a crucial role in regulating numerous physiological processes, including metabolism, neurotransmission, and hormone action. Measuring ADCY levels is particularly relevant in studies of signal transduction, endocrine function, and various metabolic and cardiovascular diseases.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to ADCY, allowing capture of the enzyme from the samples.
- Standards:
- Solutions containing known concentrations of recombinant or purified ADCY, used to generate a standard curve for quantification of ADCY in unknown 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 the ADCY protein, often conjugated to an enzyme like HRP for detection.
- Enzyme Conjugate:
- In some kits, the detection antibody may be biotinylated, requiring a streptavidin-HRP conjugate for detection.
- Substrate Solution (TMB):
- Tetramethylbenzidine, a chromogenic substrate that is converted by HRP into a colored product measurable by absorbance.
Assay Procedure Summary
- Sample and Standard Addition: Samples and standards are added to the microplate wells, where ADCY binds to the immobilized antibody.
- Wash: Unbound components are removed by washing the wells.
- Detection Antibody Addition: A detection antibody specific to another epitope of ADCY is added, binding to the captured enzyme.
- Wash: Excess detection antibody is washed away.
- Enzyme Conjugate Addition: (If applicable) A streptavidin-HRP conjugate is added, which binds to the biotinylated detection antibody.
- Wash: Remove unbound enzyme conjugate by washing.
Performance Characteristics
- Sensitivity: The assay can detect ADCY at low picogram per milliliter (pg/mL) concentrations, making it suitable for measuring low-abundance targets.
- Precision: The assay typically exhibits low intra-assay and inter-assay variability, with coefficients of variation (CV) generally below 10%.
- Linearity: The assay demonstrates linearity across the standard curve range, allowing for accurate quantification across a range of ADCY concentrations.
- Recovery: High recovery rates, usually above 80%, ensuring reliable performance across different sample types.
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