For research use only. Not for use in diagnostic procedures.
Protein expression changes of individual cytochrome P450 (CYP450) isoforms can be directly measured using the CYP450 Protein Assay – Human Induction Kits and an LC/mass spectrometry workflow for high specificity, sensitivity, and accuracy. These kits have been developed to employ multiple reaction monitoring (MRM) analysis to quantify seven CYP450 isoforms that are key proteins for induction studies: 1A2, 2B6, 2C9, 2C19, 2E1, 3A4 and 3A5. Kits with varying numbers of isoforms are available: 1) the four-isoform kit (1A2, 2B6, 3A4 and 3A5), and 2) the seven-isoform kit (1A2, 2B6, 2C9, 2C19, 2E1, 3A4 and 3A5).
- Directly measure CYP450 induction at the protein level without expensive antibodies
- Measure individual CYP450 isoforms with high specificity, sensitivity, and accuracy
- Easy-to-use kits and methodology employ a common MRM workflow
- Results are equivalent to traditional mRNA or activity assays
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Everything you need to perform protein quantitation: The starter kit contains peptide standards, sample preparation reagents and buffers, trypsin, a column, data acquisition methods, and processing methods and templates for performing protein quantitation using multiple reaction monitoring (MRM) analysis on an SCIEX Triple Quad or QTRAP System. The peptide standards for each protein are synthesized using isotopically-enriched amino acids (lysine and arginine) to provide the highest quantitative robustness. The LC/MS/MS method allows for a direct measurement of CYP450 protein levels without the use of antibodies, providing savings in both time and expense (Figure 1).
Advanced data analysis software
The data generated for CYP450 analysis is processed with MultiQuant Software. The software automatically integrates the MRM data and computes area ratios to the added peptide internal standards for all samples. Processed area ratio results are imported into a Microsoft Excel template for computation of protein level data. Results are plotted in a simple bar chart to compare sample expression levels of each CYP450 isoform to negative control expression levels.
Assay type comparison
Correlation between enzymatic activity, mRNA expression, and protein expression (as measured by LC/MS/MS) is excellent (Figure 2). Both baseline and inducible levels of the CYP450 isoforms are easily detected by the LC-MRM protein quantitation method.
Kits |
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Part Number | Kit Name/Description |
| 4445250 | Trypsin TPCK (500 μg) A modified (TPCK treated) serine protease that specifically hydrolyzes peptide bonds at the carboxyl side of lysine and arginine residues. Modified trypsin is treated with N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) to inactivate extraneous chymotryptic activity. For use with the P450 Human Induction 100 Assay Kit. Ten tubes are included in each package and each tube will be sufficient for at least 10 samples (1 mg of protein digested with a 1:20 ratio = 50 μg). |
| 4445251 | Peptide C18 Column A reversed phase C18 column specifically designed for the analysis of CYP450 isoforms by LC/MS/MS. Good for at least 200 analyses. |
| 4445252 | P450 Human Induction 100 Assay Kit The P450 Human Induction 100 Assay Kit provides a complete protocol, instrument data acquisition methods, processing methods and stable isotope labeled synthetic peptide standards for the analysis of four human CYP450 isoforms (1A2, 2B6, 3A4, and 3A5) for induction studies using LC/MS/MS on an SCIEX QTRAP® or Triple Quad™ system. Kit contains enough reagents for 100 Assays. Must order Trypsin TPCK (500 ug) and Peptide C18 Column sold separately. |
| 4445494 | P450 Human Induction Starter Kit The P450 Human Induction Starter Kit provides a complete package for the analysis of four human CYP450 isoforms (1A2, 2B6, 3A4, and 3A5) using MRM analysis on an SCIEX QTRAP or Triple Quad system. The kit contains a detailed protocol, instrument data acquisition methods, processing methods, LC column, stable isotope labeled synthetic peptide standards, trypsin, and all buffers and reagents for 100 assays. |
| 4465863 | CYP450 Peptide Standards Extended Panel Human Induction 100 Assay Kit The SCIEX CYP450 Peptide Standards Extended Panel Human Induction 100 Assay Kit provides a complete protocol, instrument data acquisition methods, processing methods and stable isotope labeled synthetic peptide standards for the analysis of seven human CYP450 isoforms (1A2, 2B6, 2C9, 2C19, 2E1, 3A4 and 3A5) for induction studies using LC/MS/MS on an SCIEX QTRAP or Triple Quad system. Kit contains enough reagents for 100 Assays. Trypsin TPCK (500 μg) and Peptide C18 Column sold separately. |
| 4445247 | Protein Preparation Kit 100 Assay Protein preparation kit containing enough reagent for the solubilization, reduction, and alkylation of 100 1-mg protein samples. The kit contains 1 vial of denaturant, 1 vial of reducing reagent, 1 vial of cysteine blocking reagent, and 6 vials of digestion buffer. Trypsin (4445250) sold separately. Note: Not for use with amine labeling reagents - for assays with amine labeling reagents use 4381664. |
| 4466004 | CYP450 Peptide Standards Extended Panel Human Induction Starter Kit The SCIEX CYP450 Peptide Standards Extended Panel Human Induction Starter Kit provides a complete package for the analysis of seven human CYP450 isoforms (1A2, 2B6, 2C9, 2C19, 2E1, 3A4 and 3A5) using MRM analysis on an SCIEX QTRAP or Triple Quad system. The kit contains a detailed protocol, instrument data acquisition methods, processing methods, LC column, stable isotope labeled synthetic peptide standards, trypsin, and all buffers and reagents for 100 assays. |