Nadph nadp assay kit

Manufactured by Promega

Sourced in United States

The NADPH/NADP+ Assay Kit is a laboratory equipment that measures the levels of NADPH and NADP+ in biological samples. It provides a quantitative determination of these important cofactors, which play crucial roles in cellular metabolism and redox reactions.

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Lab products found in correlation

Gsh gssg assay kit, by Promega (2 mentions) Gsh glo glutathione assay, by Promega (1 mentions) Cm h2dcfda, by BD (1 mentions) Dcfh da, by Beyotime (1 mentions) Gsh gssg assay kit, by Beyotime (1 mentions) Bodipy 581 591 c11, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

NADPH (2 citations) NADP/NADPH-Glo™ Assays kit (2 citations)

5 protocols using nadph nadp assay kit

Quantitative NADP/NADPH Assay

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The NADP/NADPH analysis were formed using a NADP/NADPH assay kit (Promega) following manufacturer’s instruction. Cells were collected and resuspended in 1 mL cold PBS. Then, counted cells (2 × 10⁶ cells) were resuspended in 60 μL Lysis Buffer and incubated at room temperature for 15 minutes. Lysate was then centrifuged at 1500 rpm for 5 minutes and use supernatant for the assay. The 12.5 μL of NADPH/NADP Extraction Solution was added into the NADPH/NADP sample wells and the mix solution was incubated at room temperature for 10–15 minutes. Then, 12.5 μL NADP/NADPH Extraction Solution was added to neutralize NADPH/NADP extracts for 10–15 minutes. 37.5 μL NADP/NADPH reaction mixture was then added to the mix and incubated at RT for 45 min. Monitor fluorescence intensity (Ex/Em = 540/590 nm) by SpectraMax i3 (MD).

Yuan Y., Wang C., Zhuang X., Lin S., Luo M., Deng W., Zhou J., Liu L., Mao L., Peng W., Chen J., Wang Q., Shu Y., Xue Y, & Huang P. (2022). PIM1 promotes hepatic conversion by suppressing reprogramming-induced ferroptosis and cell cycle arrest. Nature Communications, 13, 5237.

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Quantifying NADPH and GSH in FASN Cells

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Total cell lysates were used to determine NADPH levels in FASN-WT/-KO cells in 6 cm dish. The quantification of NADPH was assessed in total cell lysates using the NADP⁺/NADPH Assay Kit (Promega) following the manufacturer’s instructions, and intracellular NADPH was normalized to their corresponding protein content. 1×10⁴ of FASN-WT/-KO cells were seeded in 96-well plates overnight. The media was removed, and the total cellular GSH of each cell line was assessed using the GSH-Glo Glutathione Assay (Promega) following the manufacturer’s instructions, and intracellular GSH was normalized to their corresponding protein content.

Lough M., Cleland J., Langan J., Cowley A, & Wade A. (1998). Initiating angiotensin converting enzyme inhibitors in mild to moderate heart failure in general practice: randomised, placebo controlled trial. BMJ : British Medical Journal, 317(7169), 1352-1353.

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Quantifying Cellular Redox Status

Cited 2 times

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The levels of mitochondria and cellular ROS were determined by flow cytometer using CMH₂XRos (#M7513) and CM-H₂DCF-DA (#C6827) as fluorescent probes (BD Biosciences, San Jose, CA), respectively, as described in previous publications (10 (link),14 (link)). The intracellular levels of GSH/oxidized glutathione (GSSG) and NADPH/NADP+ were measured with a GSH/GSSG Assay kit (#V6612) and a NADPH/NADP+ Assay kit (#G9081) (Promega, WI) according to the manufacturer’s instructions.

Ju H.Q., Lu Y.X., Chen D.L., Zuo Z.X., Liu Z.X., Wu Q.N., Mo H.Y., Wang Z.X., Wang D.S., Pu H.Y., Zeng Z.L., Li B., Xie D., Huang P., Hung M.C., Chiao P.J, & Xu R.H. (2018). Modulation of Redox Homeostasis by Inhibition of MTHFD2 in Colorectal Cancer: Mechanisms and Therapeutic Implications. JNCI Journal of the National Cancer Institute, 111(6), 584-596.

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Measuring Cellular Redox Homeostasis

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The intracellular NADPH/NADP⁺ and GSH/GSSG ratios were measured with NADPH/NADP⁺ Assay Kit (Promega, USA, #G9081) and GSH/GSSG Assay Kit (Beyotime, Shanghai, China, S0053), respectively, according to the manufacturer’s instructions. The levels of cellular ROS were determined by flow cytometry using DCFH-DA (Beyotime, Shanghai, China, S0033s) as fluorescent probes following the manufacturer’s protocol.

Guan J., Li M., Wang Y., Zhang Y., Que Y., Lu S., Wang J., Zhu J., Huang J., Zhen Z., Sun F., Song M, & Zhang Y. (2024). MTHFD1 regulates the NADPH redox homeostasis in MYCN-amplified neuroblastoma. Cell Death & Disease, 15(2), 124.

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Quantifying Cellular Redox Homeostasis

Cited 1 time

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Cellular ROS levels were determined by flow cytometry using a CM-H2DCF-DA (#D399) Assay kit (Thermo Fisher Scientific, U.S.A.), as described in previous publications [16 (link)]. Lipid ROS levels were measured by flow cytometry using BODIPY 581/591 C11 (#D3861, Thermo Fisher Scientific, U.S.A.) staining. The intracellular levels of GSH/oxidized glutathione (GSSG) and NADPH/NADP⁺ were measured using a GSH/GSSG Assay kit (#V6612) and an NADPH/NADP⁺ Assay kit (#G9081) (Promega, WI) according to the manufacturer’s instructions.

Mo H.Y., Wang R.B., Ma M.Y., Zhang Y., Li X.Y., Wen W.R., Han Y, & Tian T. (2024). MTHFD2-mediated redox homeostasis promotes gastric cancer progression under hypoxic conditions. Redox Report : Communications in Free Radical Research, 29(1), 2345455.

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