Standard Curve (Sample)
Phosphoenolpyruvate Assay Kit
Phosphoenolpyruvate Assay Kit
Synonyms
Phosphoenolpyruvate; N/A; Phosphoenolpyruvate Assay Kit; Phosphoenolpyruvate assay kit
Detection Method
Fluorometric
Preparation and Storage
Upon receipt, store Pyruvate Kinase and Pyruvate Oxidase at -80°C. Store the 10X Assay Buffer and the MgCl2 at room temperature. Store the rest of the components at -20ºC. The Fluorometric Probe is light sensitive and must be stored accordingly. Avoid multiple freeze/thaw cycles.
Standard Curve (Sample)
Standard Curve (Sample)
Related Product Information for Phosphoenolpyruvate assay kit
Background: Phosphoenolpyruvate (PEP) is an important biochemical intermediate. The phosphate bond of PEP has the highest energy found in living organisms, and the bond plays an important role in metabolic pathways such as glycolysis, where pyruvate kinase generates ATP to form pyruvate. PEP also plays a role in gluconeogenesis, where it is formed from the decarboxylation of oxaloacetate catalyzed by phosphoenolpyruvate carboxykinase. In plants, PEP is used to make various aromatic compounds, and is involved in the process of converting inorganic carbon to organic carbon (carbon fixation). In bacteria, PEP is the source of energy for the phosphotransferase system. The Phosphoenolpyruvate Assay Kit is a simple fluorometric assay that measures the amount of phosphoenolpyruvate present in biological samples in a 96-well microtiter plate format. Each kit provides sufficient reagents to perform up to 100 assays*, including blanks, PEP standards, and unknown samples. Sample PEP concentrations are determined by comparison with a known PEP standard. The kit has a detection sensitivity limit of 3.1 uM PEP.
Principle of the Assay: The Phosphoenolpyruvate Assay Kit measures total PEP within biological samples. PEP in the presence of ADP is converted by Pyruvate Kinase to pyruvate + adenosine triphosphate (ATP). Pyruvate is converted by pyruvate oxidase in the presence of phosphate and oxygen into acetyl phosphate, carbon dioxide, and hydrogen peroxide. The resulting hydrogen peroxide is then detected with a highly specific fluorometric probe. Horseradish peroxidase catalyzes the reaction between the probe and hydrogen peroxide, which bind in a 1:1 ratio. Samples are compared to a known concentration of PEP standard within the 96-well microtiter plate format. Samples and standards are incubated for 30 minutes and then read with a standard 96-well fluorometric plate reader (Figure 1).
Principle of the Assay: The Phosphoenolpyruvate Assay Kit measures total PEP within biological samples. PEP in the presence of ADP is converted by Pyruvate Kinase to pyruvate + adenosine triphosphate (ATP). Pyruvate is converted by pyruvate oxidase in the presence of phosphate and oxygen into acetyl phosphate, carbon dioxide, and hydrogen peroxide. The resulting hydrogen peroxide is then detected with a highly specific fluorometric probe. Horseradish peroxidase catalyzes the reaction between the probe and hydrogen peroxide, which bind in a 1:1 ratio. Samples are compared to a known concentration of PEP standard within the 96-well microtiter plate format. Samples and standards are incubated for 30 minutes and then read with a standard 96-well fluorometric plate reader (Figure 1).
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Product Notes
The Phosphoenolpyruvate (Catalog #AAA44691) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Phosphoenolpyruvate, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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