SKU: AFG-BV-0190

AffiASSAY® Cathepsin B Activity Fluorometric Assay Kit

Vendor AffiGEN
Regular price €386,00 EUR
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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.