SKU: AFG-BV-0312

AffiASSAY® GAPDH Colorimetric Activity Assay Kit

Vendor AffiGEN
Regular price €172,00 EUR
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AffiASSAY® GAPDH Colorimetric Activity Assay Kit

The AffiASSAY® GAPDH Colorimetric Activity Assay Kit provides a reliable method for the quantification of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) activity in various biological samples. This assay kit offers a colorimetric detection method, enabling researchers to study glycolysis, cellular metabolism, and GAPDH-associated pathways with precision and convenience.

Key Features:

  • High Sensitivity: The assay kit can detect low levels of GAPDH activity, ensuring accurate quantification in biological samples.
  • Wide Application: The AffiASSAY® GAPDH Colorimetric Activity Assay Kit is compatible with various sample types, including cell lysates, tissues, and purified enzyme preparations.
  • 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 GAPDH activity, reducing experimental costs without compromising data quality.

Assay Principle:

The assay kit measures GAPDH activity based on the enzymatic conversion of glyceraldehyde 3-phosphate (G3P) to 1,3-bisphosphoglycerate (1,3-BPG), coupled with the reduction of NAD+ to NADH. The increase in absorbance at 450 nm is directly proportional to the GAPDH activity in the sample.

Applications:

  • Glycolysis Studies: Investigate the role of GAPDH in the glycolytic pathway and energy metabolism.
  • Cellular Metabolism Research: Study the regulation of GAPDH activity in response to metabolic changes and cellular signaling pathways.
  • Drug Discovery: Screen for compounds that modulate GAPDH activity as potential therapeutics for metabolic disorders and cancer.

Kit Contents:

  • GAPDH Assay Buffer
  • GAPDH Substrate Mix
  • GAPDH Enzyme Mix
  • NAD+
  • 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® GAPDH Colorimetric Activity 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® GAPDH Colorimetric Activity Assay Kit
  • Biological samples containing GAPDH
  • Microplate reader capable of measuring absorbance at 450 nm
  • Pipettes and tips
  • Microcentrifuge tubes

Protocol:

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 GAPDH 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 GAPDH Assay Buffer by diluting as instructed in the kit manual.
  • Add the GAPDH Substrate Mix and GAPDH Enzyme Mix 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 reaction to proceed.

Colorimetric Measurement:

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

Data Analysis:

  • Calculate the GAPDH activity 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 GAPDH 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® GAPDH Colorimetric Activity 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.