SKU: AFG-BV-0342

AffiASSAY® Glutamine Colorimetric Assay Kit

Vendor AffiGEN
Regular price 5 170 SEK
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AffiASSAY® Glutamine Colorimetric Assay Kit

The AffiASSAY® Glutamine Colorimetric Assay Kit offers a reliable method for the quantification of glutamine in various biological samples. This assay kit provides a colorimetric detection method, enabling researchers to study glutamine metabolism, amino acid metabolism, and cellular signaling pathways with precision and convenience.

Key Features:

  • High Sensitivity: The assay kit can detect low levels of glutamine, ensuring accurate quantification in biological samples.
  • Wide Application: The AffiASSAY® Glutamine Colorimetric Assay Kit is compatible with various sample types, including cell lysates, tissue homogenates, and culture media.
  • 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 glutamine levels, reducing experimental costs without compromising data quality.

Assay Principle:

The assay kit measures glutamine based on the enzymatic conversion of glutamine to glutamate by glutaminase, coupled with the oxidation of NADH to NAD+. The decrease in absorbance at 450 nm is directly proportional to the glutamine concentration in the sample.

Applications:

  • Metabolism Studies: Investigate glutamine metabolism, including its role in amino acid biosynthesis, protein synthesis, and nitrogen transport.
  • Cell Signaling: Study the involvement of glutamine in cellular signaling pathways, including mTOR signaling and glutaminolysis.
  • Cancer Research: Explore the role of glutamine metabolism in cancer cell proliferation, survival, and metastasis.

Kit Contents:

  • Glutamine Assay Buffer
  • Glutaminase Enzyme Mix
  • NADH
  • Microplate (96 wells)
  • Glutamine Standard
  • 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® Glutamine 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® Glutamine Colorimetric Assay Kit
  • Biological samples containing glutamine
  • Microplate reader capable of measuring absorbance at 450 nm
  • Pipettes and tips
  • Microcentrifuge tubes

Protocol:

Standard Curve Preparation:

  • Prepare a series of standard solutions containing known concentrations of the glutamine standard provided in the kit.
  • Dilute the Glutamine Assay Buffer and Glutaminase Enzyme Mix 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 glutamine 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 Glutamine Assay Buffer, Glutaminase Enzyme Mix, and NADH.
  • 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 enzymatic reaction to proceed.

Colorimetric Measurement:

  • After the incubation period, measure the absorbance at 450 nm using a microplate reader.

Data Analysis:

  • Calculate the glutamine concentrations in your samples based on the absorbance readings and 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 glutamine 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® Glutamine 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.