NAD(P)H-Glo™ Detection System
Monitor the Concentration of Reduced Forms of NADH and NADPH
- Highly sensitive and rapid bioluminescent assay
- Easily adapted for high-throughput format
- Simple add-mix-measure protocol
Catalog Number:
Size
Catalog Number: G9061
Catalog Number: G9062
The NAD(P)H-Glo™ Detection System is a bioluminescent, homogeneous, single-reagent-addition assay that quantitatively monitors the concentration of the reduced forms of NADH and NADPH levels in enzymatic reactions. The oxidized forms, NAD+ and NADP+, are not detected and do not interfere with quantitation.
The assay is rapid, requiring only a 40- to 60-minute incubation, has a broad linear range (25nM-50µM) and high signal-to background ratio (~400). Low nanomolar sensitivity, 25nM, allows direct in-well detection. The assay is well suited to measuring NAD(P)H production or consumption in high-throughput formats.
Assay Chemistry
In the presence of NAD(P)H, a reductase reduces the proluciferin reductase substrate to form luciferin. Luciferin is quantified using Ultra-Glo™ Recombinant Luciferase, which is a component of the Luciferin Detection Reagent, and the light signal produced is proportional to the amount of NAD(P)H in the sample.
Sensível e Seletivo
O ensaio é seletivo para as formas reduzidas dos níveis de NADH e NADPH em reações enzimáticas.
Compatible with High-Throughput Systems
Os protocolos simples de adição e leitura são compatíveis com o fluxo de trabalho automatizado e de alto rendimento, e os produtos químicos flexíveis se adaptam aos formatos de placas de 96, 384 e 1.536 poços e aos sistemas de alto rendimento. Eles produzem valores Z′ >0,7.
Os ensaios foram realizados usando 8 µl da concentração indicada de NADH com 8 µl do Reagente de Detecção de Luciferina apropriado em placas de 384 poços. S/B = relação sinal/plano de fundo.
The NAD(P)H-Glo™ Assay detects only NADH and NADPH.
Estudo de Caso
Os pesquisadores da BioAscent implementaram o ensaio NAD(P)H-Glo™ para HTS. Nesse projeto, o Dr. McElroy e sua equipe usaram o Ensaio NAD(P)H-Glo™ para examinar 50.000 compostos contra uma enzima mal caracterizada que é um alvo potencial de medicamentos. O NAD(P)H-Glo foi escolhido por ter alta sensibilidade, com uma ampla faixa dinâmica e altas proporções de sinal para fundo.
Protocols
Complete Protocol
Specifications
Catalog Number:
What's in the box?
| Item | Código | Tamanho | Concentration |
|---|---|---|---|
Reductase |
G884A | 1 × 55μl | 6mg/ml |
Reductase Substrate |
G885A | 1 × 55μl | |
|
Luciferin Detection Reagent |
V859A | 1 × 1 each | |
|
Reconstitution Buffer |
V865A | 1 × 10ml |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
What's in the box?
| Item | Código | Tamanho | Concentration |
|---|---|---|---|
Reductase |
G884B | 1 × 275μl | 6mg/ml |
Reductase Substrate |
G885B | 1 × 275μl | |
|
Luciferin Detection Reagent |
V859B | 1 × 1 each | |
|
Reconstitution Buffer |
V865B | 1 × 50ml |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
Resources
Citations
-
Secreted APE1/Ref-1 inhibits TNF-α-stimulated endothelial inflammation via thiol-disulfide exchange in TNF receptor.
2016
Sci. Rep.
-- -
Large-scale label-free comparative proteomics analysis of polo-like kinase 1 inhibition via the small-molecule inhibitor BI 6727 (Volasertib) in BRAFV600E mutant melanoma cells.
2014
J. Proteome Res.
-- -
Crystal structure of the emerging cancer target MTHFD2 in complex with a substrate-based inhibitor.
2017
Cancer Res.
-- -
Compensation for intracellular environment in expression levels of mammalian circadian clock genes.
2014
Sci. Rep.
-- - See all citations
Related Products
Produtos semelhantes
Nicotinamide Adenine Dinucleotide Cell-Based Assays
Sensitive, bioluminescent assays to detect NAD(P)+ and NAD(P)H levels in cells.
G9071, G9072, G9081, G9082
Glucose Uptake-Glo™ Assay
Ensaio não radioativo para medir a captação de glicose.
J1341, J1342, J1343
Lactate-Glo™ Assay
Ensaio biolomunescente que detecta rapidamente o lactato em uma variedade de tipos de amostras.
J5021, J5022
GSH/GSSG-Glo™ Assay
Homogeneous assay to quantify total glutathione and glutathione ratios.
V6611, V6612
Frequentemente usado com
RealTime-Glo™ MT Cell Viability Assay
A bioluminescent method to kinetically monitor viability in cell culture up to 72 hours.
G9711, G9712, G9713
GloMax® Discover System
High-performance microplate reader for detecting luminescence, fluorescence and absorbance.
GM3000
GloMax® Navigator System
Dedicated microplate luminometer.
GM2000, GM2010
