NADP-Malic Enzyme (NADP-ME) Activity Colorimetric Assay Kit (E-BC-K555-M)
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For research use only.
| Detection Principle |
Malic enzyme (ME) is widely present in eukaryotes and prokaryotes and participates in multiple metabolic pathways. Its main metabolic substrate is malic acid, which mainly catalyzes the oxidation and decarboxylation of malic acid to pyruvate and CO2, accompanied by the reduction of NAD(P)+ to NAD(P)H. Malic acid is an important intermediate product in the tricarboxylic acid cycle. Under the catalysis of ME, malic acid is converted into pyruvate and NAD(P)H to regulate cellular energy, REDOX balance and the synthesis of biological macromolecules. In mammalian cells, malic enzymes mainly consist of NADP+ dependent ME1 in the cytoplasm, NAD+ dependent ME2 in the mitochondria, and NADP+ dependent ME3 in the mitochondria. Inhibiting ME1 can induce cellular senescence or apoptosis to suppress the growth of tumor cells. The detection principle of this kit: The enzyme catalyzes the substrate reaction to generate NADPH, which has a characteristic absorption peak at 340 nm. The activity of NADP-ME can be determined by measuring the rate of increase in absorbance at 340 nm. |
| Sample Type | Animal tissue,Plant tissue,Cell |
| Detection Method | Colorimetric method |
| Detection Instrument | Microplate reader(340 nm) |
| Research Area | Tricarboxylic Acid（TCA）Cycle,Mitochondria Function,Energy Metabolism |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid Period | 12 months |
| Sensitivity | 2.68 U/L |
| Detection Range | 2.68-338.96 U/L |
| Precision | inter-assay CV: 1.5%-2.7% | intra-assay CV: 1.3%-1.4% |
| Sample Volume | 20 μL(tissue homogenate/cell homogenate) |
| Assay Time | 30 min |
The recommended dilution factor for different samples is as follows (for reference only):
| Sample Type | Dilution Factor |
|---|---|
| 10% Mouse liver tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 10% Mouse heart tissue homogenate | 1 |
| 10% Mouse brain tissue homogenate | 1 |
| 10% Mouse spleen tissue homogenate | 1 |
| 10% Mouse lung tissue homogenate | 1 |
| 10% Mouse muscle tissue homogenate | 1 |
| 10% Potato tissue homogenate | 1 |
| 10% Corn tissue homogenate | 1 |
| 10% Cauliflower tissue homogenate | 1 |
| 1×10^6 RAW 264.7 cells | 1 |
| 1×10^6 A549 cells | 1 |
| 1×10^6 4T1 cells | 1 |
The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
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