CheKine™ Pyruvate Dehydrogenase (PDH) Activity Assay Kit (Colorimetric) | KTB1270

CheKine™ Pyruvate Dehydrogenase (PDH) Activity Assay Kit (Colorimetric) | KTB1270 | Gentaur UK, US & Europe Distribution

Product Category: Cytology

Application: Cell Metabolism

Product Type: Biochemical Assay Kit

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Kit Component: • DCFH-DA (10 mM)

Features & Benefits : • provides a simple, sensitive, rapid ROS detection method.
• based on the fluorescent probe DCFH-DA .
• suitable for flow cytometry or fluorescence microscopy.
• Provides detailed sample preparation and results calculation methods.

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Usage Note: • Do not mix the components between different batch numbers and manufacturers; otherwise, the results may be abnormal.
• Avoid bubbles while mixing or redissolving components.
• Change pipette tips frequently to avoid cross contamination between components.
• Ensure that all components and equipment are at the proper temperature before starting the experiment.
• The working concentration of positive control H2O2 should be 100 μM (Recommend during 50-200 μM depend on the cell types ) . The level of Reactive Oxygen Species (ROS) will increase significantly after stimulated 30 min-4 h. The level of Reactive Oxygen Species (ROS) maybe very different for different cells while stimulating with the positive control. The stimulating time and the concentration of positive control could be prolonged or increased, if there is no obvious rise after 30 min . Or shorten the time or reduce the concentration of positive control, while the fluorescent signal is too strong.
• During the experiment, the fluorescent probe DCFH-DA can be diluted in 1: 2000-1: 5000, and make the final concentration of DCFH-DA to 2-5 μM if the signal of negative control also too strong. The time of probe incubating can be adjusted in 15-60 minutes, and minimize the exposure time while reading the film, and the exposure time of the control group and test groups should to be consistent.
• Reactive Oxygen Species (ROS) positive control (H2O2) is only used as a sample as a positive control, and not all test groups need positive control.
• Please wash away the unbound prode after incubating the prode, otherwise it will cause a high background.
• Try to shorten the detection time after incubating the probe (except for stimulation time) to reduce various possible errors.

Storage Conditions: Please store the whole kit at -20°C upon receipt, and protect from light. The kit is valid for 12 months. Please refer to the table below Materials supplied and Storage conditions to store all the components.

Shipping: Gel pack with blue ice.

Background: Reactive Oxygen Species, Ros is natural by-products of oxygen normal metabolism, play an important role in cell signaling and in vivo balance. However, in the oxidative stress correlation state, the ROS level can be greatly increased. The accumulation of ROS will seriously damage the cell structure. Oxidative stress plays an important role in the research of cardiovascular disease, diabetes, osteoporosis, stroke, inflammatory disease, and neurodegenerative diseases and cancer, etc. ROS detection will help determine how oxidative stress adjusts various intracellular pathways.

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Tag: Reactive Oxygen Species ，ROS