Description
Mouse CYP2E1 (Cytochrome P450, family 2, subfamily E, polypeptide 1) ELISA Kit
The Mouse CYP2E1 (Cytochrome P450 Family 2 Subfamily E Polypeptide 1) ELISA Kit is specifically designed for the precise measurement of CYP2E1 levels in mouse serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring accurate and reproducible results, making it an invaluable tool for a variety of research applications.CYP2E1 is a key enzyme in the cytochrome P450 family, playing a crucial role in drug metabolism, detoxification processes, and the activation of various compounds.
Dysregulation of CYP2E1 has been implicated in a range of diseases, including liver diseases, cancer, and alcohol-induced toxicity, making it an important target for investigation and potential therapeutic interventions.With its advanced technology and reliable performance, the Mouse CYP2E1 ELISA Kit provides researchers with a powerful tool to study CYP2E1 levels in biological samples, furthering our understanding of its function and potential implications in various physiological and pathological processes.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 3.13-200 ng/mL |
| Sensitivity: | 1.88 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse CYP2E1 in samples. No significant cross-reactivity or interference between Mouse CYP2E1 and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse CYP2E1. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse CYP2E1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse CYP2E1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse CYP2E1. The concentration of Mouse CYP2E1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | CYP2E1: Metabolizes several precarcinogens, drugs, and solvents to reactive metabolites. Inactivates a number of drugs and xenobiotics and also bioactivates many xenobiotic substrates to their hepatotoxic or carcinogenic forms. Belongs to the cytochrome P450 family. |
| UniProt Protein Details: | Protein type:Oxidoreductase; Lipid Metabolism - arachidonic acid; Lipid Metabolism - linoleic acid; Endoplasmic reticulum; EC 1. 14. 13. n7; Xenobiotic Metabolism - drug metabolism - cytochrome P450; Mitochondrial; Xenobiotic Metabolism - metabolism by cytochrome P450 Cellular Component: Golgi membrane; intrinsic to endoplasmic reticulum membrane; intracellular membrane-bound organelle; membrane; mitochondrion; endoplasmic reticulum Molecular Function:3-(3-hydroxy-phenyl)propionate hydroxylase activity; toluene-sulfonate methyl-monooxygenase activity; xylene monooxygenase activity; chlorophenol 4-monooxygenase activity; 6-hydroxy-3-methyl-2-oxo-1,2-dihydroquinoline 6-monooxygenase activity; metal ion binding; oxidoreductase activity; carbon disulfide oxygenase activity; toluene 4-monooxygenase activity; phenanthrene 9,10-monooxygenase activity; 1-hydroxy-2-oxolimonene 1,2-monooxygenase activity; 2-hydroxy-phenylacetate hydroxylase activity; 4-chlorobenzaldehyde oxidase activity; enzyme binding; 2-oxo-delta3-4,5,5-trimethylcyclopentenylacetyl-CoA 1,2-monooxygenase activity; oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; limonene monooxygenase activity; dibenzothiophene monooxygenase activity; 3,5-xylenol methylhydroxylase activity; phenanthrene 1,2-monooxygenase activity; 3-methyl-2-oxo-1,2-dihydroquinoline 6-monooxygenase activity; toluene 3-monooxygenase activity; styrene monooxygenase activity; monooxygenase activity; dimethyl sulfide monooxygenase activity; 4-hydroxyphenylacetate 3-monooxygenase activity; 4-nitrophenol 4-monooxygenase activity; 1-hydroxy-2-naphthoate hydroxylase activity; toluene-4-sulfonate monooxygenase activity; oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen; tetrahydrofuran hydroxylase activity; oxygen binding; phenylacetate hydroxylase activity; toluene 2-monooxygenase activity; iron ion binding; 6-hydroxynicotinate 3-monooxygenase activity; heme binding; arachidonic acid epoxygenase activity; phenanthrene 3,4-monooxygenase activity; alpha-pinene monooxygenase [NADH] activity; steroid hydroxylase activity; 2-octaprenyl-3-methyl-6-methoxy-1,4-benzoquinol hydroxylase Biological Process: steroid metabolic process; triacylglycerol metabolic process; xenobiotic metabolic process; exogenous drug catabolic process; monoterpenoid metabolic process; epoxygenase P450 pathway; heterocycle metabolic process; drug metabolic process |
| UniProt Code: | Q05421 |
| NCBI GenInfo Identifier: | 461828 |
| NCBI Gene ID: | 13106 |
| NCBI Accession: | Q05421. 1 |
| UniProt Secondary Accession: | Q05421,Q9Z198, |
| UniProt Related Accession: | Q05421 |
| Molecular Weight: | 56,805 Da |
| NCBI Full Name: | Cytochrome P450 2E1 |
| NCBI Synonym Full Names: | cytochrome P450, family 2, subfamily e, polypeptide 1 |
| NCBI Official Symbol: | Cyp2e1 |
| NCBI Official Synonym Symbols: | Cyp2e |
| NCBI Protein Information: | cytochrome P450 2E1; CYPIIE1; cytochrome P450-J; cytochrome P450-ALC; 4-nitrophenol 2-hydroxylase; cytochrome P450, 2e1, ethanol inducible |
| UniProt Protein Name: | Cytochrome P450 2E1 |
| UniProt Synonym Protein Names: | 4-nitrophenol 2-hydroxylase (EC:1. 14. 13. n7); CYPIIE1; Cytochrome P450-ALC; Cytochrome P450-J |
| Protein Family: | Cytochrome |
| UniProt Gene Name: | Cyp2e1 |
| UniProt Entry Name: | CP2E1_MOUSE |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 200 | 2.17 2.196 | 2.183 | 2.126 |
| 100 | 1.437 1.469 | 1.453 | 1.396 |
| 50 | 0.828 0.818 | 0.823 | 0.766 |
| 25 | 0.346 0.386 | 0.366 | 0.309 |
| 12.5 | 0.204 0.18 | 0.192 | 0.135 |
| 6.25 | 0.155 0.133 | 0.144 | 0.087 |
| 3.13 | 0.103 0.103 | 0.103 | 0.046 |
| 0 | 0.051 0.063 | 0.057 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse CYP2E1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse CYP2E1 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 9.12 | 17.12 | 70.91 | 8.42 | 17.36 | 77.71 |
| Standard deviation | 0.63 | 0.77 | 2.55 | 0.57 | 0.94 | 3.42 |
| C V (%) | 6.91 | 4.50 | 3.60 | 6.77 | 5.41 | 4.40 |
Recovery
The recovery of Mouse CYP2E1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 90-102 | 97 |
| EDTA plasma (n=5) | 91-106 | 98 |
| Cell culture media (n=5) | 87-98 | 93 |
Linearity
Samples were spiked with high concentrations of Mouse CYP2E1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 93-106 | 99-115 | 94-105 |
| Average (%) | 101 | 105 | 99 | |
| 1:4 | Range (%) | 90-106 | 85-99 | 89-103 |
| Average (%) | 97 | 91 | 95 | |
| 1:8 | Range (%) | 85-97 | 79-91 | 86-100 |
| Average (%) | 92 | 85 | 91 | |
| 1:16 | Range (%) | 92-104 | 83-94 | 88-103 |
| Average (%) | 99 | 89 | 94 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
