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Enzyme Assays Kits on ZAGENO

Enzymes are specialized proteins or in some cases RNA molecules which speed up reactions in the cell. Without the presence of enzymes, chemical reactions would occur at a much slower pace - so much so life could not be sustained. Enzymes work by binding to the active site of their particular substrate, lower the reactions activation energy, thereby increasing the rate of reaction. This reaction rate is simply defined as the speed in which the reaction occurs, and is influenced by a number of factors such as temperature, pressure or concentration.

Given an enzyme’s ability to catalyze a reaction, they are vital for cellular processes such as signaling pathways, metabolism, and gene expression. Thus in knowing enzyme activity within the cell, i.e. the moles of substrate converted per unit time, scientists can infer the mechanism behind reactions. Enzyme assays are then standardized laboratory procedures which detect the level of enzymes present in a sample.

Detection Technique

So how are enzymes detected? By measuring the disappearance of the substrate, or the appearance of the product. This can be done in a variety of ways, but the most common are done either through colorimetry, fluorescence, or luminescence.

- Spectrophotometry measures the change of light absorption in the solution. Colorimetry is a type of spectrophotometric assay specific for measuring visible light.

- Fluorometric assays are similar to spectrophotometric assays in that they measure light. However, fluorescence measures light emittance after absorbing light at a different wavelength. So fluorometric assays measure the difference in fluorescence from the substrate to the product to measure the enzyme reaction.

- Luminescence measures the emission of light via chemical reaction. During the enzymatic reactions, light emittance can be used to infer product formation.

Enzyme Assays

Some standard enzyme assays measure the activity of kinase, phosphatase, superoxide dismutase, and Beta-Galactosidase.

Kinases are classed as phosphotransferases as they catalyze the transfer of phosphate groups from high-energy molecules to substrates. This process is known as phosphorylation (where a substrate gains a phosphate), and is a key process of glycolysis, the metabolic pathway common to most cells. Thus, kinases are essential for analyzing metabolic activity in cells.

Directly opposite of kinases are the phosphatases, which removes phosphate groups from a substrate.

Superoxide Dismutase catalyzes the partitioning of the superoxide ( O2- ) into ordinary 02 or H2O2. The superoxide molecule is a by-product of oxygen metabolism and can cause cell damage if not regulated. Thus superoxide dismutase is a major defensive mechanism within cells.

Beta-Galactosidase enzymes catalyze the breakdown of Beta-galactosides into monosaccharides, providing energy and free carbon to the cell.

Compare Enzyme Assay Kits!

With our compare function, you can avoid all the time and energy wasted sifting through multiple web pages from different suppliers. At ZAGENO you can clearly see kits side-by-side, with the relevant attributes for each kit neatly in line for easy selection of the best product for you.

Click on the comparison below for a clearer view!

This example highlights how every kit has positive and negative aspects, and that only side-by-side comparison can allow you to correctly judge which is the right kit for you.

For instance, this comparison shows:

  • that the ADP-Glo™ Max Assay kit is the cheapest per preperation.
  • that the Pyruvate Kinase Activity Assay Kit has controls included.
  • that the PIP2:PS Lipid Kinase Substrate uses fluorescence as detection technique and can accept tissue samples.

The ZAGENO comparison engine does not show the best kit, but instead helps you, the researcher, choose the most sutiable kit for your experiment.

Go to our How it Works page for a guide to using the ZAGENO comparison engine.

Expand your understanding in our Knowledge Section. Perfect your experiments with our Troubleshooting Guides. Join in the discussion within our Community. Learn about ZAGENO at How It Works.

Video Credits:

Video: Bozeman Science/Youtube

Discussion

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JD Watson


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