AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit
AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit is designed to provide precise and reliable measurements of cathepsin B enzyme activity in biological samples. This assay kit offers a fluorometric detection method, making it a valuable tool for researchers in the fields of biochemistry, cell biology, and cancer research.
Key Features:
- High Sensitivity: Our assay kit is optimized to detect cathepsin B activity, allowing for accurate quantification in a wide range of samples.
- Fluorometric Detection: Utilizing a highly sensitive fluorometric assay, our kit ensures robust and reproducible results, minimizing the margin for error.
- Wide Application: The AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit can be used with various sample types, including cell lysates, tissue homogenates, serum, plasma, and culture supernatants.
- Quick and Easy: With a simple and streamlined protocol, you can obtain results in just a few easy steps, saving you valuable time in the lab.
- Versatile: Whether you're studying cathepsin B in cellular processes, investigating its role in cancer progression, or screening potential inhibitors, our kit provides the versatility you need.
Assay Principle:
This assay kit measures cathepsin B activity based on the enzymatic cleavage of a fluorogenic substrate by cathepsin B. The substrate contains a fluorophore that is quenched in its intact state but becomes highly fluorescent upon cleavage by cathepsin B, allowing for the quantification of enzyme activity.
Applications:
- Cancer Research: Assess cathepsin B activity as a biomarker for tumor progression, metastasis, and therapeutic response in various cancer types.
- Cell Biology: Study the role of cathepsin B in intracellular protein degradation, autophagy, and lysosomal function.
- Drug Discovery: Screen compounds for their effects on cathepsin B activity and evaluate their potential as therapeutic agents for diseases associated with cathepsin dysregulation.
Kit Contents:
- Assay Buffer
- Fluorogenic Substrate
- Cathepsin B Enzyme
- Stop Solution
- 96-Well Assay Plate
- Detailed Protocol
Why Choose AffiASSAY®?
Our commitment to excellence and scientific rigor makes AffiASSAY® the trusted choice of researchers worldwide. We strive to provide you with innovative tools that deliver precise results and accelerate your research efforts.
AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit Protocol:
[Please note that this protocol is a general guideline, and you should always refer to the specific instructions provided in the kit manual for accurate and detailed information.]
Materials Needed:
- AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit
- Biological samples containing cathepsin B
- 96-well microplate
- Fluorescence microplate reader capable of measuring fluorescence intensity at specific wavelengths
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Standard Curve Preparation:
- Reconstitute the Cathepsin B Enzyme provided in the kit according to the kit's instructions.
- Create a series of standard dilutions using the reconstituted Cathepsin B Enzyme to generate a standard curve. Typical concentrations may range from 0 to 1000 U/mL.
Assay Plate Setup:
- Label the microplate wells according to your experimental design, including wells for your samples, standards, and controls.
- Prepare control wells containing assay buffer only as background controls.
Sample and Standard Addition:
- Add 50-100 µL of your prepared samples and Cathepsin B Standards to the appropriate wells on the microplate.
- Ensure that the final reaction volume in each well is consistent.
Assay Reaction:
- Add 50 µL of Fluorogenic Substrate to each well to initiate the reaction.
- Incubate the microplate at the appropriate temperature (as specified in the kit instructions) for a recommended time to allow the enzymatic cleavage of the substrate.
Reaction Termination:
- Add 50 µL of Stop Solution to each well to stop the reaction.
Fluorescence Measurement:
- Measure the fluorescence intensity of each well using a fluorescence microplate reader at the excitation and emission wavelengths provided by the kit.
Data Analysis:
- Generate a standard curve using the fluorescence values obtained from the Cathepsin B Standards.
- Calculate the cathepsin B activity in your samples based on their respective fluorescence readings and the standard curve.
Results Interpretation:
- Interpret and analyze your results according to your research objectives.
- Note any variations or trends in cathepsin B activity among your samples.
Data Presentation:
- Prepare your data for presentation or publication, and include the appropriate statistical analysis if required.
Cleanup:
- Dispose of all waste materials, including used assay components, according to your laboratory's safety protocols.
- Always refer to the specific instructions provided with the AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit for any kit-specific details or variations in the protocol.
- Additionally, ensure that you adhere to the best practices and safety guidelines in your laboratory while conducting the assay.