The ATF (Activating Transcription Factor) ELISA Kit is designed for the quantitative measurement of ATF proteins in various biological samples, including cell lysates, tissue homogenates, and nuclear extracts. ATF proteins belong to the ATF/CREB family of transcription factors, which play critical roles in the regulation of gene expression in response to cellular stress, growth factors, and various signaling pathways. These factors are involved in processes such as apoptosis, metabolism, and development, making them important targets for research in oncology, neurobiology, and cellular stress responses.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to ATF, enabling the capture of ATF proteins from the sample.
- Standards:
- Calibrated solutions containing known concentrations of recombinant or purified ATF, used to generate a standard curve for quantifying ATF 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 ATF protein, often conjugated to an enzyme such as HRP for detection.
- Enzyme Conjugate:
- In some kits, this may be a secondary antibody conjugated to HRP or a biotinylated detection antibody with a streptavidin-HRP conjugate.
- Substrate Solution (TMB):
- Tetramethylbenzidine, which is converted by HRP to a colored product that can be measured spectrophotometrically.
Assay Procedure Summary
- Sample and Standard Addition: Add standards and samples to the microplate wells where ATF proteins are captured by the immobilized antibody.
- Wash: Unbound components are removed by washing the wells.
- Detection Antibody Addition: A detection antibody specific to another epitope of ATF is added, which binds to the captured protein.
- Wash: Excess detection antibody is washed away.
- Enzyme Conjugate Addition: (If applicable) An HRP-conjugated enzyme or a streptavidin-HRP conjugate is added to bind the detection antibody.
- Wash: Remove unbound enzyme conjugate through washing.
Performance Characteristics
- Sensitivity: The assay can detect ATF proteins at low picogram per milliliter (pg/mL) concentrations, making it suitable for detecting low-abundance targets.
- Precision: The assay typically 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 across different concentrations of ATF.
- Recovery: High recovery rates, usually above 85%, ensuring accurate and reliable performance across various sample types.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|