The intracellular levels of pyruvate, α-KG, and acetyl-CoA were determined by using commercial kits (Biovision). In brief, 2×106 cells were homogenized in PBS. The supernatant was collected, and the proteins were removed by using 10 KD Amicon Ultra Centrifugal Filters (Millipore). The flow-through containing the metabolites was used for the measurement of pyruvate, α-KG, and acetyl-CoA, following the manufacturer’s instructions. Intracellular GSH/GSSG ratio was determined using GSH/GSSG-Glo™ (Promega), following the manufacturer’s instructions. Total cellular ATP levels in live cells were assessed using an ATP assay kit (Sigma Aldrich).
Kd amicon ultra centrifugal filters
Manufactured by Merck Group
The KD Amicon Ultra Centrifugal Filters are a type of laboratory equipment used for the purification and concentration of macromolecules, such as proteins and nucleic acids, through the process of tangential flow filtration. The filters are designed to efficiently separate and retain target analytes from complex solutions, allowing for the removal of unwanted substances and the concentration of the desired molecules.
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Lab products found in correlation
3 protocols using kd amicon ultra centrifugal filters
1
Intracellular Metabolite Quantification
2
Quantification of TCA Cycle Metabolites
3
Intracellular Metabolite Quantification
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The intracellular levels of pyruvate, α-KG, and acetyl-CoA were determined by using commercial kits (Biovision). In brief, 2×106 cells were homogenized in PBS. The supernatant was collected, and the proteins were removed by using 10 KD Amicon Ultra Centrifugal Filters (Millipore). The flow-through containing the metabolites was used for the measurement of pyruvate, α-KG, and acetyl-CoA, following the manufacturer’s instructions. Intracellular GSH/GSSG ratio was determined using GSH/GSSG-Glo™ (Promega), following the manufacturer’s instructions. Total cellular ATP levels in live cells were assessed using an ATP assay kit (Sigma Aldrich).
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