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Mouse CYP1A2(Cytochrome P450 1A2)ELISA Kit (MOES01781)

SKU:
MOES01781
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P00186
Sensitivity:
0.09ng/mL
Range:
0.16-10ng/mL
ELISA Type:
Sandwich
Reactivity:
Mouse
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Metabolism
€649
Frequently bought together:

Description

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Mouse CYP1A2 (Cytochrome P450 1A2) ELISA Kit

The Mouse CYP1A2 (Cytochrome P450 1A2) ELISA Kit is a valuable tool for measuring levels of CYP1A2 in mouse serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring accurate and consistent results for various research applications.CYP1A2 is a key enzyme involved in the metabolism of drugs, toxins, and endogenous compounds in the liver.

It plays a crucial role in drug metabolism and activation, making it a significant target for studying drug interactions, toxicity, and pharmacokinetics.By utilizing the Mouse CYP1A2 ELISA Kit, researchers can better understand the role of CYP1A2 in various physiological and pathological processes, leading to potential advancements in drug development and personalized medicine.

Assay type:Sandwich
Format:96T
Assay time:4.5h
Reactivity:Mouse
Detection Method:Colormetric
Detection Range:0.16-10 ng/mL
Sensitivity:0.10 ng/mL
Sample Volume Required Per Well:100µL
Sample Type:Serum, plasma and other biological fluids
Specificity:This kit recognizes Mouse CYP1A2 in samples. No significant cross-reactivity or interference between Mouse CYP1A2 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 CYP1A2. 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 CYP1A2 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 CYP1A2, 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 CYP1A2. The concentration of Mouse CYP1A2 in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:CYP1A2: Cytochromes P450 are a group of heme-thiolate monooxygenases. In liver microsomes, this enzyme is involved in an NADPH-dependent electron transport pathway. It oxidizes a variety of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics. Most active in catalyzing 2-hydroxylation. Caffeine is metabolized primarily by cytochrome CYP1A2 in the liver through an initial N3-demethylation. Also acts in the metabolism of aflatoxin B1 and acetaminophen. Participates in the bioactivation of carcinogenic aromatic and heterocyclic amines. Catalizes the N-hydroxylation of heterocyclic amines and the O- deethylation of phenacetin. Belongs to the cytochrome P450 family. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Secondary Metabolites Metabolism - caffeine; Xenobiotic Metabolism - metabolism by cytochrome P450; Oxidoreductase; Amino Acid Metabolism - tryptophan; EC 1. 14. 14. 1; Xenobiotic Metabolism - drug metabolism - cytochrome P450; Cofactor and Vitamin Metabolism - retinol; Lipid Metabolism - linoleic acid

Cellular Component: endoplasmic reticulum; intracellular membrane-bound organelle; membrane

Molecular Function:demethylase activity; enzyme binding; heme binding; iron ion binding; metal ion binding; monooxygenase activity; oxidoreductase activity; oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; 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

Biological Process: alkaloid metabolic process; aromatic compound metabolic process; cellular respiration; dibenzo-p-dioxin metabolic process; drug catabolic process; drug metabolic process; exogenous drug catabolic process; heterocycle metabolic process; hydrogen peroxide biosynthetic process; lipid metabolic process; lung development; monocarboxylic acid metabolic process; monoterpenoid metabolic process; porphyrin metabolic process; post-embryonic development; regulation of gene expression; steroid catabolic process; steroid metabolic process; toxin biosynthetic process; toxin metabolic process

UniProt Code:P00186
NCBI GenInfo Identifier:6753566
NCBI Gene ID:13077
NCBI Accession:NP_034123. 1
UniProt Secondary Accession:P00186,Q9QWJ4,
UniProt Related Accession:P00186
Molecular Weight:58,184 Da
NCBI Full Name:cytochrome P450 1A2
NCBI Synonym Full Names:cytochrome P450, family 1, subfamily a, polypeptide 2
NCBI Official Symbol:Cyp1a2
NCBI Official Synonym Symbols:CP12; Cyp1a1; P450-3
NCBI Protein Information:cytochrome P450 1A2
UniProt Protein Name:Cytochrome P450 1A2
UniProt Synonym Protein Names:CYPIA2; Cholesterol 25-hydroxylase
Protein Family:Cytochrome
UniProt Gene Name:Cyp1a2
UniProt Entry Name:CP1A2_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.DAverageCorrected
102.285
2.301		2.2932.228
51.564
1.616		1.591.525
2.50.879
0.855		0.8670.802
1.250.375
0.411		0.3930.328
0.630.256
0.23		0.2430.178
0.320.166
0.152		0.1590.094
0.160.106
0.122		0.1140.049
00.055
0.075		0.065--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse CYP1A2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse CYP1A2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.501.483.890.541.563.58
Standard deviation0.030.070.200.030.080.12
C V (%)6.004.735.145.565.133.35

Recovery

The recovery of Mouse CYP1A2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5)83-9690
EDTA plasma (n=5)96-112102
Cell culture media (n=5)93-10899

Linearity

Samples were spiked with high concentrations of Mouse CYP1A2 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)
1:2Range (%)87-10287-10085-100
Average (%)949391
1:4Range (%)93-10586-9684-97
Average (%)1009190
1:8Range (%)94-10785-9784-94
Average (%)1009189
1:16Range (%)94-10884-9689-103
Average (%)1009094

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
  1. 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.
  2. Aliquot 100µl of standard solutions into the standard wells.
  3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
  4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
  5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
  6. 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.
  7. 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.
  8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
  9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
  10. 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.
  11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
  12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

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