The DNM ELISA Kit is used for the quantitative measurement of DNM (Dynamin) proteins in biological samples. Dynamin is a large GTPase protein involved in various cellular processes, including endocytosis, vesicle trafficking, and membrane remodeling. It plays a critical role in the pinching off of vesicles from the plasma membrane and is essential for various cellular functions.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to Dynamin, allowing for capture of Dynamin from the samples.
- Standards:
- Solutions containing known concentrations of recombinant or purified Dynamin, used to generate a standard curve for quantifying Dynamin 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 Dynamin, 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
- Coating:
- Coat the wells of the microplate with an antibody specific to Dynamin. Incubate to allow binding and then wash away unbound antibody.
- 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 Dynamin in the samples will bind to the immobilized antibody.
- Incubation:
- Incubate the plate to allow Dynamin to bind to the antibody.
- Addition of Detection Antibody:
- Add a detection antibody specific to Dynamin, which binds to the captured Dynamin.
Performance Characteristics
- Sensitivity: Capable of detecting low levels of Dynamin, 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
- Cellular Trafficking Research: Studying the role of Dynamin in endocytosis, vesicle budding, and membrane dynamics.
- Neurological Studies: Investigating Dynamin’s role in synaptic vesicle recycling and neurodegenerative diseases.
- Cancer Research: Evaluating Dynamin levels in cancer cells and understanding its role in tumor progression.
- Drug Development: Assessing the impact of potential therapeutics on Dynamin function and cellular trafficking.
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