In the DCFH‐DA assay, a DCFH fluorescence probe (Beyotime, Nantong, China) was diluted into a concentration of 10 μL/10 mL using DMEM without FBS. In dark conditions, PBS washed cells were subjected to incubation with a diluted DCFH fluorescence probe (1 mL/well) for 30 minutes in a cell incubator without light, then digested with trypsin and resuspended in DMEM containing FBS for neutralizing trypsin. Cells were washed with PBS twice before detection by flow cytometric analysis at emission wavelengths of 519 nm. In the NAPDH assay, NADPH content was detected using an NADP/NADPH Kit (Biovision, Milpitas, CA, USA) according to the manufacturer's instructions.
Nadp nadph kit
Manufactured by Abcam
Sourced in United States
The NADP/NADPH Kit is a colorimetric assay kit designed to quantify the levels of NADP (nicotinamide adenine dinucleotide phosphate) and NADPH (the reduced form of NADP) in various sample types. The kit provides a simple and reliable method for the detection and measurement of these important cofactors.
Automatically generated - may contain errors
Lab products found in correlation
Nad nadh kit, by Abcam (1 mentions)
Cobas 8000, by Roche (1 mentions)
Dihydroethidium, by Merck Group (1 mentions)
Px 478, by Selleck Chemicals (1 mentions)
Agi 6780, by Selleck Chemicals (1 mentions)
Octyl α ketoglutarate, by Cayman Chemical (1 mentions)
β action antibody, by Merck Group (1 mentions)
Cocl2, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
6 protocols using nadp nadph kit
1
Oxidative Stress Evaluation via DCFH-DA and NADPH
2
Quantifying NADP+/NADPH in HCAEC
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HCAEC (3×105 per well) were incubated (1 h, 37°C, 5% CO2) in HBSS with 0 or 500 µM MGO. After washing twice with HBSS, and freeze-thawing in extraction buffer, total (NADP++NADPH), and NADPH alone, were quantified using a NADP+/NADPH kit (BioVision, Mountainview, CA, USA), according to the manufacturers’ instructions.
3
Measurement of Cellular Redox Cofactors
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Production of NADH and NADPH was measured using, respectively, the NAD/NADH kit (Abcam, Cambridge, UK) and the NADP/NADPH kit (Abcam) according to the manufacturer's instruction. Lactate and LDH activity were measured using the Roche diagnostic kit on a Cobas 8000 (Roche Diagnostics, Mannheim, DE, USA) as described previously.39 (link)
4
Metabolic Profiling of Cellular Responses
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CoCl2, dihydroethidium and propidium iodide were from Sigma/Aldrich. Octyl-α-ketoglutarate was from Cayman and PX-478 and AGI-6780 were from Selleck. Antibodies to detect IDH2, HIF1α, LDHA, ALDOA, PDK1, and total ubiquitination and the NADP/NADPH kit were from Abcam and the β-action antibody was from Sigma.
5
Determination of Adenine Nucleotides
6
NADPH/NADP Ratio Measurement
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Cells were cultured and extracted using NADPH/NADP extraction buffer. The NADPH/NADP ratio was measured using commercial NADP+/NADPH kits (Abcam).
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