A new proteomics technology by LC-MS/MS/MRM Quantitative proteomics tools & analysis service by iMPAQT method
Date：October 11 2019Web Page No：81385
We offer product & service to quantify multiple proteins at one time by mass spectrometer (LC-MS/MS)
- iMPAQT*1, New technology for Quantitative Proteomics
- Technology Outline
- Standard peptides mixture for LC-MS/MS MRM- 338x Human major metabolic enzymes can be quantified-MRMplus Standard Mix
- Workflow: Preparation of Scheduled MRM by MRMplus®
- Contract service
iMPAQT*1, New technology for Quantitative Proteomics
＊1 in vitro proteome-assisted MRM for Protein Absolute QuanTification
Matsumoto, M., et al., Nat. Methods, 14（3）, 251～258（2017）.［PMID: 28267743］
- Developed by Drs. Nakayama & Matsumoto of Kyushu University.
- Highly ionized peptides by actual measurement of genome-wide human recombinant proteins
- Quantitative profiling of ~400 proteins/hour at once
- A powerful tool for pathway-wide proteomics, subtype clarification and no antibodies available targets
Technology Outline
- In vitro synthesis of 18,000 recombinant human proteins, followed by digestion to peptides & mTRAQ*2 (heavy) labeling.
- Acquire pre-information (MRM transition parameters, ionization values, relative retention time) of every peptides by LC-MS/MS.
- MRM-measurement based on the pre-information for the samples & internal standards labeled with mTRAQ*2 (light for samples, heavy for standards)
＊2 mTRAQ reagent is product of AB Sciex
Standard peptides mixture for LC-MS/MS MRM- 338x Human major metabolic enzymes can be quantified-MRMplus Standard Mix
Features
Total 338 of major human metabolic enzymes from carbohydrate metabolism, lipid metabolism, amino acids metabolism, nucleic acid metabolism.
Only 2 days !
(Sample prep.: 1 day + MS analysis: 1 day) Expression level of 338x metabolic enzymes in one profile.
Expression level of human major metabolic enzymes in cultured cells
*This product is licensed from Kyushu university.
*This product requires triple-quadrupole (QqQ) mass spectrometry (AB Sciex 4000QTRAP or later).
*mTRAQ reagent (AB Sciex) is required but not provided.
*Compatible with conventional LC: Column inner diameter should be less than 2.1mm.
Other Features
- Simple: Well-validated 338 human major metabolic enzymes & 8 endogenous controls are pre-mixed, just mTRAQ-label & Go!
- Easy: Pre-set parameters (MRM transition, relative retention time) will be provided.
Application
- Visualization of up/down-regulations on whole metabolic pathway.
- Drug discovery/Pharmacology
- Basic research for proteomics
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[Date : February 15 2024 12:37]
Related Products: Proteomics Sample Preparation Kit for Cultured Cell 2G
Cell lysis & peptide preparation for mass spectrometry by cultured cell sample, optimized for mTRAQ *1 conjugation for LC-MS/MS MRM.
*1: mTRAQ is a product from AB Sciex
Protocol overview
- Cell lysis
- BCA assay*2
- Protease digestion & alkylation
- mTRAQ *2 Labeling.
- Freeze-drying (-65oC).
*2: BCA assay kit and mTRAQ reagents are not provided but required.
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Proteomics Sample Preparation Kit for Cultured Cell 2G |
FMR-003 | FMRFunakoshi Co.,Ltd. | 1 kit | $380 | |||||||||||||||||||||||||||||||
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Related Products: MRMplus Retention Time Marker
A mixture of 12 synthetic peptides for calibration of retention times for each peptides in LC-MS/MS system. Compatible with MRMplus Standard Mix.
The result of LC-MS/MS MRM by this product
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MRMplus Retention Time Marker DatasheetThis may not be the latest data sheet. |
FMR-002 | FMRFunakoshi Co.,Ltd. | 12 µl | $150 | |||||||||||||||||||||||||||||||
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MRMplus Retention Time Marker
DatasheetThis may not be the latest data sheet.
- Product Code: FMR-002
- Supplier: FMR
- Size: 12µl
- Price: $150
| Description |
MRMplus Retention Time Marker is the mixture of marker peptides (12 synthetic peptides) for calculating the retention times of LC in LC-MS/MS system for scheduled-MRM. The segment value of each MRMplus Standard Peptide is easily converted by using this marker and the conversion software. |
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| Storage | -80°C | CAS | |
| Link | |||
Workflow: Preparation of Scheduled MRM by MRMplus®
MRM (Multiple Reaction Monitoring) and is one of the quantitative proteomics methods by mass spectrometry.
MRM uses triple-quadrupole LC-MS/MS and enable us to distinguish peptides by their amino acid sequences specifically by detecting product/fragment ions of selected MS parameters (MRM transitions).
It is possible to quantify proteins of interest (POIs) by measuring MRM transitions of specific sequences (Proteotypic Peptides: PTPs) in samples along with internal standard peptides.
Contract service
Proteomics specialists offer protein quantitative analysis of metabolic pathway-wide proteome profiling using the latest MS instrument & MRM Standard Mix.
- Available panel：Human major metabolic enzyme (338 proteins)
- Sample preps：Frozen pellets of human cultured cells or frozen human tissue
※Sample must be sent with serum free condition.
Example #1: Quantitative Analysis of Metabolic Enzymes in 11 Human Cancer Cell Lines
Bottom: Quantitative analysis was performed against 266 x human major metabolic enzymes in 11 human cancer cells. The expression levels were shown in the heat map below. Distinguished expression patterns among cell lines, which are clustered by the profile similarities. iMPAQT enables to show important factors on each pathway by calculating enzyme expression as quantitative value.
Right: Bubble charts of expression levels for glycolysis enzymes in HeLa cells. Expression levels of upstream enzymes were low, while those of downstream ones were high.
Example #2: Profiles of Human Metabolic Enzymes among Cell Cycles
After synchronizing cell division with thymidine block, HeLa cells were collected at every 2 hours and 236 x metabolic enzyme expressions were measured.
In S phase, expressions of many metabolic enzyme are suppressed (Green). In G1 phase, the expression level related to glycolysis and TCA cycle were greatly increased (Red).
NOTE: This service uses synthetic peptides as standard, so obtained quantitative value is standardized by synthetic peptides (It does not reflect protein purification at pre-treatment and efficiency of enzyme digestion). Obtained quantitative value is calculated from concentration of inner standard peptide.
CONTACT
export@funakoshi.co.jp
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- ※Please note that Product Information or Price may change without notice.