SKU: AFG-BV-0398

AffiASSAY® Hydroxyproline Colorimetric Assay Kit

Vendor AffiGEN
Regular price $655.00 AUD
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AffiASSAY® Hydroxyproline Colorimetric Assay Kit

The AffiASSAY® Hydroxyproline Colorimetric Assay Kit provides a reliable method for the quantification of hydroxyproline levels in various biological samples. Hydroxyproline is a major component of collagen, and its measurement is crucial for studying collagen turnover, extracellular matrix remodeling, and tissue fibrosis. This assay kit offers a sensitive colorimetric detection method, allowing researchers to accurately assess hydroxyproline content with ease.

Key Features:

  • High Sensitivity: The assay kit can detect low levels of hydroxyproline, ensuring accurate quantification in biological samples with minimal interference.
  • Wide Application: Compatible with various sample types, including tissues, cell lysates, and biological fluids, the AffiASSAY® Hydroxyproline Colorimetric Assay Kit offers versatility in collagen-related research.
  • Quick and Easy: With a streamlined protocol, you can obtain results rapidly, saving time and resources in the laboratory.
  • Cost-Effective: Our kit provides an affordable solution for measuring hydroxyproline levels, reducing experimental costs without compromising data quality.

Assay Principle:

The assay kit measures hydroxyproline levels based on the chloramine-T oxidation method. Hydroxyproline reacts with chloramine-T to form an intermediate, which reacts with p-dimethylaminobenzaldehyde (DMAB) to produce a colored compound. The intensity of the color is directly proportional to the concentration of hydroxyproline in the sample.

Applications:

  • Collagen Turnover: Study collagen synthesis and degradation processes in physiological and pathological conditions.
  • Extracellular Matrix Remodeling: Assess tissue remodeling events associated with wound healing, fibrosis, and tissue repair.
  • Fibrosis Research: Investigate the role of hydroxyproline in tissue fibrosis and explore potential therapeutic interventions.

Kit Contents:

  • Hydroxyproline Assay Buffer
  • Hydroxyproline Chloramine-T Reagent
  • Hydroxyproline DMAB Reagent
  • Hydroxyproline Standard
  • Microplate (96 wells)
  • 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® Hydroxyproline Colorimetric 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® Hydroxyproline Colorimetric Assay Kit
  • Biological samples containing hydroxyproline
  • Microplate reader capable of measuring absorbance
  • Pipettes and tips
  • Microcentrifuge tubes

Protocol:

Standard Curve Preparation:

  • Prepare a series of standard solutions containing known concentrations of hydroxyproline.
  • Dilute the Hydroxyproline Assay Buffer, Hydroxyproline Chloramine-T Reagent, and Hydroxyproline DMAB Reagent as instructed in the kit manual.
  • Add the standard solutions to each well of the microplate to generate the standard curve.

Assay Plate Setup:

  • Label the microplate wells according to your experimental design, including wells for your samples, standards, and controls.
  • If using controls, prepare control samples without hydroxyproline to serve as background controls.

Sample and Standard Addition:

  • Add 50-100 µL of your prepared samples and standard solutions to the appropriate wells on the microplate.
  • Ensure that the final reaction volume in each well is consistent.

Assay Reaction:

  • Prepare the reaction mixture by combining the Hydroxyproline Assay Buffer, Hydroxyproline Chloramine-T Reagent, and Hydroxyproline DMAB Reagent.
  • Add the reaction mixture to each well of the microplate containing the samples and standards.
  • Incubate the microplate at the appropriate temperature (as specified in the kit instructions) for a recommended time to allow the color development.

Colorimetric Measurement:

  • After the incubation period, measure the absorbance at the appropriate wavelength using a microplate reader.

Data Analysis:

  • Calculate the hydroxyproline levels in your samples based on the absorbance readings and the standard curve generated from the standard solutions.

Results Interpretation:

  • Interpret and analyze your results according to your research objectives.
  • Observe any variations or trends in hydroxyproline levels 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® Hydroxyproline Colorimetric 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.