NAD+ is a redox cofactor important to the right functioning of a wide range of essential metabolic pathways, together with key steps in mitochondrial vitality metabolism. As well as, it serves as a signaling substrate for enzymes reminiscent of sirtuins and the poly-ADP ribosyl-polymerase household of enzymes.
Sirtuins, that are NAD+-dependent protein deacylases, harness modifications in mobile NAD+ concentrations to provide modifications in protein acylation standing, thereby affecting downstream features together with vitality metabolism, stress resistance, and cell survival.
Thus, the provision of NAD+ in cells, or in particular organelles such because the mitochondrion, regulates downstream signaling and key organic processes. This idea has pushed a necessity for researchers to simply and exactly measure NAD+ concentrations in organic samples.
We herein describe a number of protocols for the measurement of NAD+ and NADH concentrations in tissues, cells, or subcellular compartments reminiscent of mitochondria. These protocols embody a biking assay that may shortly measure NAD+ or NADH ranges utilizing a plate reader outfitted with fluorescence measurement capabilities.
This plate assay depends solely upon commercially out there supplies along with the organic samples of curiosity. As well as, we describe a protocol using steady isotope-labeled NAD+ as an inside customary to find out organic NAD+ content material by isotope-dilution strategies. This technique requires mass spectrometry to ratio endogenous NAD+ with exogenous isotope-labeled NAD+ to acquire quantification utilizing HPLC and mass spectrometry.
MULTIPLEX KIT PCR MASTITIS PCR kit |
PCR-MPX218-48D |
Bioingentech |
MULTIPLEX KIT PCR MASTITIS PCR kit |
PCR-MPX218-96D |
Bioingentech |
MULTIPLEX KIT PCR Babesia & Theileria PCR kit |
PCR-MPX401-48D |
Bioingentech |
MULTIPLEX KIT PCR Babesia & Theileria PCR kit |
PCR-MPX401-96D |
Bioingentech |
Blood PCR Kit |
20-abx09801320ulSystems |
Abbexa |
A novel light-controlled colorimetric detection assay for nitroreductase primarily based on p-aminophenol-catalyzed and NADH-mediated synthesis of silver nanoparticles
A novel and environment friendly light-controlled colorimetric assay for the quantification and detection of nitroreductase (NTR) was constructed primarily based on p-aminophenol (pAP)-catalyzed and nicotinamide adenine dinucleotide (NADH)-mediated technology of AgNPs.
Because of the hydrolysis of p-nitrophenol by NTR within the presence of NADH, the hydrolysis product can be utilized as a catalyst to catalyze the discount of Ag+ by NADH beneath the sunshine. Because the focus of NTR will increase, the worth of absorbance at ca. 400 nm (A400) decreases and the colour of the answer turns from brown to vivid yellow.
A linear correlation was obtained between A400 and the NTR focus within the vary from 1-50 μg mL-1 and the restrict of detection (LOD) is 0.27 μg mL-1.
The detection system doesn’t reply to different widespread organic molecules because of the specificity of enzymes and the impact of the nitroreductase inhibitor on the NTR exercise was additionally examined.
Lastly, we utilized the assay to find out NTR in human serum samples by spiking totally different concentrations of NTR with a restoration of 85.2%-92.5%.
Identification of proteins able to metallic discount from the proteome of the Gram-positive bacterium Desulfotomaculum reducens MI-1 utilizing an NADH-based exercise assay.
Understanding of microbial metallic discount relies nearly solely on research of Gram-negative organisms. On this research, we deal with Desulfotomaculum reducens MI-1, a Gram-positive metallic reducer whose genome lacks genes with similarity to any characterised metallic reductase.
Utilizing non-denaturing separations and mass spectrometry identification, together with a colorimetric display screen for chelated Fe(III)-NTA discount with NADH as electron donor, we’ve got recognized proteins from the D. reducens proteome not beforehand characterised as iron reductases.
Their perform was confirmed by heterologous expression in Escherichia coli. Moreover, we present that these proteins have the aptitude to scale back soluble Cr(VI) and U(VI) with NADH as electron donor. The proteins recognized are NADH : flavin oxidoreductase (Dred_2421) and a protein complicated composed of oxidoreductase flavin adenine dinucleotide/NAD(P)-binding subunit (
Paraffin Wax) and dihydroorotate dehydrogenase 1B (Dred_1686).
Dred_2421 was recognized within the soluble proteome and is predicted to be a cytoplasmic protein. Dred_1685 and Dred_1686 had been recognized in each the soluble in addition to the insoluble protein fraction, suggesting a sort of membrane affiliation, though PSORTb predicts each proteins are cytoplasmic. This research is the primary purposeful proteomic evaluation of D. reducens and one of many first analyses of metallic and radionuclide discount in an environmentally related Gram-positive bacterium.
Pir ELISA Kit| Rat Pirin ELISA Kit |
EF019186 |
Lifescience Market |
REN ELISA Kit| Rat Renin ELISA Kit |
EF017245 |
Lifescience Market |
Kl ELISA Kit| Rat Klotho ELISA Kit |
EF017474 |
Lifescience Market |
LN ELISA Kit| Rat Laminin ELISA Kit |
EF017041 |
Lifescience Market |
Lep ELISA Kit| Rat Leptin ELISA Kit |
EF017074 |
Lifescience Market |
NADH-driven poly-3-hydroxybutyrate accumulation in Escherichia coli: Knowledge from enzymatic assays and oxygen-limited steady cultures
Biosynthesis of poly-3-hydroxybutyrate (PHB) as a fermentation product allows the coupling of progress and product technology. Furthermore, the discount of oxygen provide ought to cut back operative value and enhance product yield.
Era of PHB as a fermentation product relies on the in vivo exercise of an NADH-preferring acetoacetyl-CoA reductase.
Proof of this idea requires
(i) quantification of the cofactor desire, in physiologically related circumstances, of a putative NADH-preferring acetoacetyl-CoA reductase and
(ii) verification of PHB accumulation utilizing an NADH-preferring acetoacetyl-CoA reductase in a species naturally incapable of doing so, for instance, Escherichia coli.
This dataset comprises kinetic knowledge obtained by spectrophotometry and knowledge from a steady tradition of an engineered E. coli pressure accumulating PHB beneath oxygen-limiting circumstances.
On this dataset it’s attainable to search out
(1) enzyme stability assays;
(2) preliminary charges and progress curves from reactions catalyzed by two acetoacetyl-CoA reductases;
(3) estimations of the relative use of NADH and NADPH by two acetoacetyl-CoA reductases;
(4) estimations of the flux capability of the response catalyzed by an acetoacetyl-CoA reductase;
(5) biomass composition of an engineered E. coli pressure remodeled with a plasmid;
(6) calculation of reconciled particular charges of this engineered pressure rising on sucrose as the only real carbon supply beneath oxygen limitation and
(7) metabolic fluxes distributions throughout the steady progress of this engineered pressure.
As a result of a comparatively small variety of acetoacetyl-CoA reductases have been kinetically characterised, knowledge and scripts right here offered may very well be helpful for additional kinetic characterizations.
Furthermore, the process described to estimate biomass composition may very well be fascinating to estimate plasmid and protein burden in different strains. Software of knowledge reconciliation to fermentations ought to assist to acquire particular charges in step with the precept of mass and electron conservation.
All of the required knowledge and scripts to carry out these analyses are deposited in a Mendeley Knowledge repository. This text was co-submitted with the manuscript entitled “An NADH preferring acetoacetyl-CoA reductase is engaged in poly-3-hydroxybutyrate accumulation in Escherichiasia. coli”.
The next sensitivity and effectivity of widespread primer multiplex PCR assay in identification of meat origin utilizing NADH dehydrogenase subunit Four gene.
A Widespread Primer Multiplex PCR (CP-M-PCR) was developed to detect meat origin of 4 teams of animal (pig, ruminant, avian and rabbit). This technique demonstrated increased sensitivity and effectivity than the traditional multiplex PCR.
On this method, a standard ahead primer was designed within the 5′ finish of a homologous area of mitochondrial NADH dehyrogenase subunit 4 (Nad 4) gene sequences of all of the animal teams. Particular adapter reverse primers had been designed by including an adapter sequence on the 5′ finish.
The identical adapter sequence was used because the widespread adapter reverse primer. The primers generated particular fragments of 267, 370, 504, and 548 bp lengths for pig, ruminant, avian and rabbit meats, respectively. The usage of adapter sequence on the 5′ finish of the widespread adapter reverse primers elevated the effectivity of the amplification and the appliance of a standard ahead primer solved the complexity in multiplex PCR system.
Bands of particular amplification will be detected within the PCR assays containing as little as 10(-6) μM of adapter reverse primer. This end result indicated that the sensitivity was tremendously elevated as in comparison with the traditional multiplex PCR (10(-3) μM).
EnzyChrom™ NAD/NADH Assay Kit |
E2ND-100 |
BioAssay Systems |
100 |
EUR 479 |
Description: For sensitive determination of NAD and NADH and evaluation of drug effects on NAD/NADH metabolism. Key Features: Sensitive and accurate. Detection limit of 0.05 µM and linearity up to 10 µM NAD+/NADH in 96-well plate assay. Convenient. The procedure involves adding a single working reagent, and reading the optical density at time zero and 15 min at room temperature. High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Method: OD565nm. Samples: Cell or tissue extracts. Species: All. Procedure: Assay takes 15 min. Kit size: 100 tests. Detection limit: 0.05 µM. |
Amplite® Colorimetric NADH Assay Kit |
15271 |
AAT Bioquest |
400 Tests |
EUR 278 |
Amplite® Colorimetric NADH Assay Kit |
15271-400Tests |
AAT Bioquest |
400 Tests |
EUR 278 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
PicoProbe? NADH Fluorometric Assay Kit |
K338-100 |
Biovision |
each |
EUR 692.4 |
Amplite® Fluorimetric NADH Assay Kit *Red Fluorescence* |
15261 |
AAT Bioquest |
400 Tests |
EUR 334 |
Amplite® Colorimetric Total NAD and NADH Assay Kit |
15258 |
AAT Bioquest |
400 Tests |
EUR 334 |
Amplite® Fluorimetric NADH Assay Kit *Red Fluorescence* |
15261-400Tests |
AAT Bioquest |
400 Tests |
EUR 334 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
Amplite® Colorimetric Total NAD and NADH Assay Kit |
15258-400Tests |
AAT Bioquest |
400 Tests |
EUR 334 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
Amplite® Colorimetric NAD/NADH Ratio Assay Kit |
15273 |
AAT Bioquest |
250 Tests |
EUR 390 |
Amplite® Colorimetric NAD/NADH Ratio Assay Kit |
15273-250Tests |
AAT Bioquest |
250 Tests |
EUR 390 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
EZScreen? NAD+/NADH Colorimetric Assay Kit (384-well) |
K958-400 |
Biovision |
each |
EUR 777.6 |
Amplite® Fluorimetric Total NAD and NADH Assay Kit *Red Fluorescence* |
15257 |
AAT Bioquest |
400 Tests |
EUR 334 |
Amplite® Fluorimetric Total NAD and NADH Assay Kit *Red Fluorescence* |
15257-400Tests |
AAT Bioquest |
400 Tests |
EUR 334 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
Amplite® Colorimetric Total NAD and NADH Assay Kit *Enhanced Sensitivity* |
15275 |
AAT Bioquest |
400 Tests |
EUR 334 |
Amplite® Colorimetric Total NAD and NADH Assay Kit *Enhanced Sensitivity* |
15275-400Tests |
AAT Bioquest |
400 Tests |
EUR 334 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
Amplite® Fluorimetric NAD/NADH Ratio Assay Kit *Red Fluorescence* |
15263 |
AAT Bioquest |
250 Tests |
EUR 390 |
Amplite® Fluorimetric NAD/NADH Ratio Assay Kit *Red Fluorescence* |
15263-250Tests |
AAT Bioquest |
250 Tests |
EUR 390 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
NADPase Assay Kit |
abx097983-100Assays |
Abbexa |
100 Assays |
EUR 604.8 |
|
NADP / NADPH Assay Kit |
abx096008-100Assays |
Abbexa |
100 Assays |
EUR 566.4 |
|
NADP+/NADPH Assay Kit |
MET-5018 |
Cell Biolabs |
100 assays |
EUR 686.4 |
Description: Our NADP+/NADPH Assay Kit detects NADP+ and NADPH in cell and tissue lysates. Total NADP+/NADPH can be detected or samples can be treated with an acid or base treatment to specifically detect NADP+ or NADPH. This assay uses an enzymatic cycling reaction that reduces NADP+ to NADPH, which then reacts with a colorimetric probe and is detected with a spectrophotometric plate reader at 450nm. NADP+/ NADPH levels in unknown samples are calculated based on the provided NADP+ standard curve. |
NADP/NADPH Assay Kit |
Z5030038 |
Biochain |
100 assays |
EUR 805 |
NAD Kinase Assay Kit |
abx298807-100Assays |
Abbexa |
100 Assays |
EUR 661.2 |
|
EnzyFluo NADP/NADPH Assay Kit |
EFNP-100 |
BioAssay Systems |
100 |
EUR 537.6 |
Description: Sensitive determination of NADP and NADPH by fluorometric (530/585nm) method. Procedure: 30 min. Kit size: 100 tests. Detection limit: 0.01 µM. Shelf life: 6 months. Shipping: on ice; storage: -20°C. |
EnzyChrom NADP/NADPH Assay Kit |
ECNP-100 |
BioAssay Systems |
100 |
EUR 537.6 |
Description: Quantitative determination of NADP and NADPH by colorimetric (565nm) method. Procedure: 30 min. Kit size: 100 tests. Detection limit: 0.1 µM. Shelf life: 6 months. Shipping: on ice; storage: -20°C. |
MicroMolar NADPH Assay Kit |
NADPH100 |
ProFoldin |
100 assays |
EUR 221.78 |
Description: This product includes 1 ml of 10x Assay buffer, 0.1 ml of 20 x Reagent A, 0.1 ml of 20 x Reagent B, 0.5 ml of 10x Reagent C and 0.020 ml of 10 mM NADPH. |
NADP+/NADPH Assay Kit (Fluorometric) |
MET-5031 |
Cell Biolabs |
100 assays |
EUR 686.4 |
Description: Our NADP+/NADPH Assay Kit detects NADP+ and NADPH in cell and tissue lysates. Total NADP+/NADPH can be detected or samples can be treated with an acid or base treatment to specifically detect NADP+ or NADPH. This assay uses an enzymatic cycling reaction that reduces NADP+ to NADPH, which then reacts with a fluorometric probe and is detected with a fluorescence plate reader at 530-570nm excitation and 590-600nm emission. NADP+/ NADPH levels in unknown samples are calculated based on the provided NADP+ standard curve. |
EnzyFluo™ NADP/NADPH Assay Kit |
EF2NP-100 |
BioAssay Systems |
100 |
EUR 479 |
Description: For quantitative determination of NADP and NADPH and evaluation of drug effects on their metabolism (This product is a replacement for the discontinued product EFNP-100). Key Features: Sensitive and accurate. Detection limit of 0.01 µM and linearity up to 1 µM NADP+/NADPH in 96-well plate assay. Convenient. The procedure involves adding a single working reagent, and reading the fluorescence at time zero and 30 min. Room temperature assay. High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Method: FL530/585nm. Samples: Cell, tissue extracts etc. Species: All. Procedure: Assay takes 30 min. Kit size: 100 tests. Detection limit: 0.01 µM. |
EnzyChrom™ NADP/NADPH Assay Kit |
E2NP-100 |
BioAssay Systems |
100 |
EUR 479 |
Description: For quantitative determination of NADP+/NADPH concentrations and ratios in cell or tissue extracts. Key Features: Sensitive and accurate. Detection limit 0.1 µM, linearity up to 10 µM NADP+/NADPH in 96-well plate assay. Convenient. The procedure involves adding a single working reagent, and reading the optical density at time zero and 30 min at room temperature. No 37°C heater is required. High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Method: Colorimetric Determination at 565 nm. Samples: Cells, Tissue. Species: human, mouse, rat and more. Procedure: Assay takes 30 min. Kit size: 100 tests. Detection limit: 0.1 µM. |
XTT CELL VIABILITY ASSAY KIT (1000 ASSAY) |
30007 |
Biotium |
1KIT |
EUR 283.2 |
Description: Minimum order quantity: 1 unit of 1KIT |
NADH-Inhibitor |
Y4681000 |
Bio Basic |
10mg |
EUR 240.96 |
|
E. coli NAD+ Kinase Assay Kit Plus |
NADK100KE |
ProFoldin |
100 assays |
EUR 444.91 |
Description: This product includes 600 ul of 10 x assay buffer, 35 ul of 100 x NAD+, 35 ul of 100 x E. coli NAD kinase, 35 ul of 100 x ATP, 35 ul of 100 x MUK Reagent A, 35 ul of 100 x MUK Reagent B and 300 ul of 10 x fluorescence dye for 100 assays in a 384-well assay format. |
Amplite® Colorimetric NADPH Assay Kit |
15272 |
AAT Bioquest |
400 Tests |
EUR 278 |
Amplite® Colorimetric NADPH Assay Kit |
15272-400Tests |
AAT Bioquest |
400 Tests |
EUR 278 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
NADPH-Cytochrome c Reductase Assay Kit |
abx097996-100Assays |
Abbexa |
100 Assays |
EUR 698.4 |
|
E. coli NAD+ Kinase Assay Kit Plus-500 |
NADK500KE |
ProFoldin |
500 assays |
EUR 1491.11 |
Description: This product includes 2 ml of 10 x assay buffer, 170 ul of 100 x NAD+, 170 ul of 100 x E. coli NAD kinase, 170 ul of 100 x ATP, 170 ul of 100 x MUK Reagent A, 170 ul of 100 x MUK Reagent B and 1.7 ml of 10 x fluorescence dye for 500 assays in a 384-well assay format. |
Amplite® Colorimetric NADP/NADPH Ratio Assay Kit |
15274 |
AAT Bioquest |
250 Tests |
EUR 390 |
Amplite® Colorimetric NADP/NADPH Ratio Assay Kit |
15274-250Tests |
AAT Bioquest |
250 Tests |
EUR 390 |
|
Description: Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. |
NADH, disodium salt |
2735-1000 |
Biovision |
each |
EUR 202.8 |
CP-M-PCR detection restrict of the DNA samples was 0.1 ng for the 4 teams of meats. CP-M-PCR has enormously improved the sensitivity and effectivity of the PCR system for a extra dependable and correct final result than typical multiplex PCR system.
Pir ELISA Kit| Rat Pirin ELISA Kit |
EF019186 |
REN ELISA Kit| Rat Renin ELISA Kit |
EF017245 |
Kl ELISA Kit| Rat Klotho ELISA Kit |
EF017474 |