Nadp nadph glo assay

Manufactured by Promega

Sourced in United States, United Kingdom

The NADP/NADPH-Glo Assay is a luminescent-based detection system for quantifying NADP and NADPH levels in biological samples. It uses a proprietary thermostable luciferase enzyme to measure NADP and NADPH concentrations with high sensitivity and linearity.

Automatically generated - may contain errors

Lab products found in correlation

Nad nadh glo assay, by Promega (7 mentions) Gsh glo glutathione assay, by Promega (4 mentions) N 5755, by Merck Group (3 mentions) Fluostar omega, by BMG Labtech (3 mentions) Six well ultra low attachment culture plates, by Corning (2 mentions) Gsh gssg glo assay, by Promega (2 mentions) Epidermal growth factor (egf), by Merck Group (2 mentions) Basic fibroblast growth factor (bfgf), by Merck Group (2 mentions) Dmem f12 50 50 medium, by Corning (2 mentions) Infinite 200 pro, by Tecan (2 mentions) Synergy 2, by Agilent Technologies (2 mentions) Fluostar omega plate reader, by BMG Labtech (2 mentions) Bca protein assay, by Thermo Fisher Scientific (2 mentions) N 6705, by Merck Group (2 mentions) Nad nadh glo, by Promega (1 mentions) Glucose colorimetric assay kit, by Cayman Chemical (1 mentions) Acetyl coenzyme a assay kit, by Merck Group (1 mentions) Glucagon quantikine elisa kit, by R&D Systems (1 mentions) Alt gpt reagent, by Thermo Fisher Scientific (1 mentions) Ast got reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

NADP/NADPH-Glo (19 citations) NADPH-Glo assay (4 citations) NADPH (2 citations) NADP/NADPH-Glo™ Assays kit (2 citations)

62 protocols using nadp nadph glo assay

Islet NAD(P)(H) Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Batches of 20 islets were preincubated 40 min in G0.5 before incubation under various conditions for 15 min. The reaction was stopped with NaOH 0.2 N and 1% DTAB and islets were briefly sonicated. NAD⁺, NADH, NADP⁺, and NADPH were assayed with the NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays (Promega) (see Suppl. Experimental Procedures).

Santos L.R., Muller C., de Souza A.H., Takahashi H.K., Spégel P., Sweet I.R., Chae H., Mulder H, & Jonas J.C. (2017). NNT reverse mode of operation mediates glucose control of mitochondrial NADPH and glutathione redox state in mouse pancreatic β-cells. Molecular Metabolism, 6(6), 535-547.

+ Open protocol

+ Expand

Metabolic Biomarker Analysis in Liver

Check if the same lab product or an alternative is used in the 5 most similar protocols

Liver TG, serum TG and serum cholesterol were measured with Triglyceride Colorimetric Assay Kit (Cayman Chemical #10010303) and Cholesterol Fluorometric Assay Kit (Cayman Chemical #10007640), respectively, according to manufacturer protocols. Serum insulin and glucagon concentrations were determined by Mouse Insulin ELISA KIT (Invitrogen, EMMINS) and Glucagon Quantikine ELISA Kit (R&D Systems, DGCG0), respectively, following manufacturer protocols. Liver NADPH and NADP were measured by NADP/NADPH-Glo^™ Assays (Promega #G9081) according to manufacturer’s protocol. ALT and AST assays were performed with ALT(GPT) Reagent (Thermo Scientific^™, TR71121) and AST/GOT Reagent (Thermo Scientific^™, TR70121), respectively, according to the manufacturer’s protocol. ATP, Acetyl-CoA and G6P concentrations were determined by ATP assay kit (Abcam, ab83355), Acetyl-Coenzyme A Assay Kit (Sigma, MAK039) and PicoProbe^™ Glucose-6-Phosphate Fluorometric Assay Kit (Biovision, K687), respectively according to the manufacturers protocols. Glucose and lactate were measured with Glucose colorimetric assay kit (Cayman Chemical #10009582) and Lactate Colorimetric/Fluorometric Assay Kit (Biovision, K607), respectively, according to the manufacturer’s protocols.

Gu L., Zhu Y., Watari K., Lee M., Liu J., Perez S., Thai M., Mayfield J.E., Zhang B., Rocha K.C., Li F., Kim L.C., Jones A.C., Wierzbicki I.H., Liu X., Newton A.C., Kisseleva T., Lee J.H., Ying W., Gonzalez D.J., Saltiel A.R., Simon M.C, & Karin M. (2023). Fructose-1,6-bisphosphatase is a nonenzymatic safety valve that curtails AKT activation to prevent insulin hyperresponsiveness. Cell metabolism, 35(6), 1009-1021.e9.

+ Open protocol

+ Expand

NADPH and NADP Measurement Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

NADPH and NADP measurement was performed using NADP/NADPH-Glo™ Assays (Promega #G9081) according to manufacturer’s protocol. Briefly, KC6141 cells were grown on 6-well plates coated with ECM and treated as above. Then the cells were resuspended at 10⁶ cells/mL 50μl of PBS and 50μl of base solution with 1% DTAB and transferred to 96-well plates. The plate was briefly mixed to ensure homogeneity and cell lysis. 50μl of each sample was removed to an empty well for acid treatment. 25μl per well of 0.4N HCl was added into these samples. All samples were incubated for 15 minutes at 60°C and then equilibrated for 10 minutes at room temperature. 25μl of 0.5M Trizma® base was added into each well of acid-treated cells to neutralize the acid. 50μl of HCl/Trizma® solution was added to each well containing base-treated samples. Then 100μl of NADP/NADPH-Glo™ Detection Reagent was added into each well and incubated for 30 minutes at room temperature. Luminescence was recorded using a luminometer.

Su H., Yang F., Fu R., Li X., French R., Mose E., Pu X., Trinh B., Kumar A., Liu J., Antonucci L., Todoric J., Liu Y., Hu Y., Diaz-Meco M.T., Moscat J., Metallo C.M., Lowy A.M., Sun B, & Karin M. (2021). Cancer Cells Escape Autophagy Inhibition via NRF2 Induced Macropinocytosis. Cancer cell, 39(5), 678-693.e11.

+ Open protocol

+ Expand

Assessing IDH2 Enzymatic Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols

The enzymatic activity of IDH2 was assessed by measuring the NADPH/NADP+ ratio in mitochondria.^{16 (link)} Mitochondria were extracted using the Cell Mitochondria Isolation Kit (Beyotime), and the NADPH/NADP+ ratio was calculated with the NADP/NADPH-Glo™ Assays (Promega, Madison, WI, USA) according to the manufacturer’s instructions.

Chen X., Zhuo S., Xu W., Chen X., Huang D., Sun X, & Cheng Y. (2020). Isocitrate dehydrogenase 2 contributes to radiation resistance of oesophageal squamous cell carcinoma via regulating mitochondrial function and ROS/pAKT signalling. British Journal of Cancer, 123(1), 126-136.

+ Open protocol

+ Expand

Metabolic Profiling of Cellular Processes

Cited 3 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Glycolysis, glycolytic capacity (ECAR), and oxidative phosphorylation (OXPHOS) were measured with the Seahorse Extracellular Flux (XF-96) analyzer (Seahorse Bioscience, North Billerica, MA) as previously described (20 (link)). The glucose uptake assay was performed as previously described (19 (link)). Briefly, cells were transfected with siRNA and 48h after transfection, glucose uptake was determined using a fluorescently labeled glucose analog, 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose (2-NBDG; Cayman Chemical Company), according to manufacturer’s instructions. Relative fluorescence was normalized to protein concentration.
Fatty acid oxidation was assessed using a previously described method (14 (link)), with minor modifications. Briefly, cells were cultured in media containing [9,10(n)-³H] palmitic acid. ³H₂O secreted into the medium was quantified by scintillation counting and normalized to the cellular protein content in each well. To measure NADPH/NADP⁺, 48h after siRNA transfection, NADPH and NADP⁺ were measured using NADP/NADPH-Glo Assay (Promega) according to the manufacturer’s protocol.

Chiyoda T., Hart P.C., Eckert M.A., McGregor S.M., Lastra R.R., Hamamoto R., Nakamura Y., Yamada S.D., Olopade O.I., Lengyel E, & Romero I.L. (2017). Loss of BRCA1 in the cells of origin of ovarian cancer induces glycolysis: A window of opportunity for ovarian cancer chemoprevention. Cancer prevention research (Philadelphia, Pa.), 10(4), 255-266.

+ Open protocol

+ Expand

Quantifying Cellular NAD(P)+/NAD(P)H Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

Intracellular NAD(P)⁺ and NAD(P)H relative levels were determined using NAD/NADH-Glo Assay and NADP/NADPH-Glo Assay (Promega, Madison, WI, USA). For both couples, cells were lysed with base solution (Triton X-100 1% v/v in NaOH 0.2 N) and the lysate was split in two parts. To measure the oxidized forms (NAD⁺ and NADP⁺), samples were treated with HCl 0.4 N at 60 °C for 15 min and incubated at room temperature for 10 min, followed by the addition of Tris base 0.5 M and NAD/NADH or NADP/NADPH-Glo Detection Reagent. To measure the reduced forms (NADH and NADPH), samples were heated at 60 °C for 15 min and incubated at room temperature for 10 min, followed by the addition of HCl-Tris solution and NAD/NADH or NADP/NADPH-Glo Detection Reagent. The luminescent signal was recorded and NAD(P)⁺/NAD(P)H ratios and relative amounts were calculated.

Cascardo F., Anselmino N., Páez A., Labanca E., Sanchis P., Antico-Arciuch V., Navone N., Gueron G., Vázquez E, & Cotignola J. (2021). HO-1 Modulates Aerobic Glycolysis through LDH in Prostate Cancer Cells. Antioxidants, 10(6), 966.

+ Open protocol

+ Expand

NADP+/NADPH Quantification in Colon Tumors

Check if the same lab product or an alternative is used in the 5 most similar protocols

NADP⁺ and NADPH analyses from frozen colon tumor samples were performed directly in 96-well plate culture plates according to the manufacturers’ instructions (NADP/NADPH Glo Assay, Promega, Madison, WI, USA). NADP⁺ and NADPH values were normalized by protein content.

Aguilar E., Esteves P., Sancerni T., Lenoir V., Aparicio T., Bouillaud F., Dentin R., Prip-Buus C., Ricquier D., Pecqueur C., Guilmeau S, & Alves-Guerra M.C. (2019). UCP2 Deficiency Increases Colon Tumorigenesis by Promoting Lipid Synthesis and Depleting NADPH for Antioxidant Defenses. Cell Reports, 28(9), 2306-2316.e5.

+ Open protocol

+ Expand

Quantifying Cellular NADPH Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

NADPH concentration was determined using the NADP/NADPH-Glo Assay (Promega), following manufacturer instructions. NADP+ levels were determined from the standard curve.

Bidault G., Virtue S., Petkevicius K., Jolin H.E., Dugourd A., Guénantin A.C., Leggat J., Mahler-Araujo B., Lam B.Y., Ma M.K., Dale M., Carobbio S., Kaser A., Fallon P.G., Saez-Rodriguez J., McKenzie A.N, & Vidal-Puig A. (2021). SREBP1-induced fatty acid synthesis depletes macrophages antioxidant defences to promote their alternative activation. Nature metabolism, 3(9), 1150-1162.

+ Open protocol

+ Expand

Assessing Cellular Redox Status

Check if the same lab product or an alternative is used in the 5 most similar protocols

GSH-Glo Glutathione Assay and NADP/NADPH-Glo Assay (both from Promega) were used to measure the levels of GSH and NADPH/NADP⁺ (total and ratio), respectively, following the manufacturer’s instructions.

Elsaadi S., Steiro I., Abdollahi P., Vandsemb E.N., Yang R., Slørdahl T.S., Rø T.B., Menu E., Sponaas A.M, & Børset M. (2021). Targeting phosphoglycerate dehydrogenase in multiple myeloma. Experimental Hematology & Oncology, 10, 3.

+ Open protocol

+ Expand

NADP+/NADPH Ratio in Yeast Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

NADP⁺, NADPH, contents in yeast cells were assessed with NADP/NADPH-Glo Assay according to the manufacturer’s protocols (Promega). Cells from early exponential phase culture were centrifuged, washed, suspended to density of 2 × 10⁶ cells/mL in PBS buffer, and used for determination purposes. Luminescence was recorded for 3 h using TECAN Infinite 200 microplate reader (Tecan Group Ltd., Männedorf, Switzerland). The value of the blank was subtracted each time. The results were presented as NADP⁺/NADPH ratio, which showed the share of NADPH and NADP⁺ in the total NADP(H) pool.

Maslanka R, & Zadrag-Tecza R. (2020). Reproductive Potential of Yeast Cells Depends on Overall Action of Interconnected Changes in Central Carbon Metabolism, Cellular Biosynthetic Capacity, and Proteostasis. International Journal of Molecular Sciences, 21(19), 7313.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!