After transfection, the levels of glutamine, α-KG, and ATP in HuCCT1 and HuH28 cells were analyzed with a Glutamine/Glutamate Determination Kit (Sigma), α-KG Assay Kit (Sigma), or ATP Colorimetric/Fluorometric Assay Kit (Sigma).
α kg assay kit
Manufactured by Merck Group
Sourced in United States
The α-KG Assay Kit is a laboratory tool designed to quantitatively measure the levels of alpha-ketoglutarate (α-KG) in various sample types. The kit provides the necessary reagents and protocols to perform accurate and reliable α-KG analysis.
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4 protocols using α kg assay kit
1
Metabolic Profiling of Transfected Cells
2
Quantifying Cellular Metabolites: ATP and α-KG
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Following the washing step with precooled PBS, intracellular ATP content was determined using the Enhanced ATP Assay Kit (Beyotime, China) as per the manufacturer’s instructions. Cellular α-Ketoglutarate (α-KG) was quantified using the α-KG Assay Kit (Sigma-Aldrich, USA). In brief, prepared equal volumes of samples and Reaction Mixes were added to a 96-well transparent microplate and read at a wavelength of 570 nm using a multifunctional microplate reader (EnSpire) (PerkinElmer, USA). The concentration of cellular α-KG was calculated using a linear regression equation for α-KG standards.
3
Rh2E2 Impacts Acetyl-CoA and α-KG
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LLC-1 cells were treated with 80 μM Rh2E2 for 24 h. The cell lysates were then harvested for determination of acetyl-coenzyme A (Acetyl-CoA) and α-Ketoglutarate (α-KG) by Acetyl-CoA Assay Kit and α-KG Assay Kit (Sigma, MO, USA) following manufacturer's instruction. Detail methodologies were described in supplementary information .
4
Quantification of α-KG in BM-MSCs
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BM-MSCs were lysed in ice-cold α-KG assay buffer. The α-KG level was measured using an α-KG Assay Kit (Sigma Aldrich, St. Louis, MO) according to the manufacturer’s instructions.
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