At Creative Proteomics, Our R&D department focuses on optimizing all aspects of peptide quantification and improving the biochemical and functional properties of your product. All development work is designed to meet the specific needs of Creative Proteomics' business and research partners. our PRM-based peptide quantification service, you can benefit from our expertise, state-of-the-art instrumentation, and comprehensive data analysis to achieve accurate and reliable peptide quantification results for your research.
What is PRM-Based Peptide Quantification?
PRM-based peptide quantification, also known as targeted proteomics or parallel reaction monitoring (PRM), is a mass spectrometry-based method used to measure the abundance or concentration of specific peptides in a complex biological sample. It is often employed in quantitative proteomics studies to determine the relative or absolute levels of proteins or peptides in a sample. In PRM-based peptide quantification, a specific set of peptides of interest is selected for measurement based on prior knowledge or a discovery proteomics approach. These peptides are synthesized or obtained as synthetic peptide standards and spiked into the sample as internal standards. The sample is then subjected to enzymatic digestion to generate a complex peptide mixture. The digested peptides are separated by liquid chromatography (LC) and introduced into a mass spectrometer, where they undergo fragmentation. Unlike traditional data-dependent acquisition (DDA) methods, where mass spectrometers autonomously select precursor ions for fragmentation, PRM uses a targeted approach. In PRM, the precursor ions of the selected peptide targets are sequentially isolated in the mass spectrometer and fragmented using higher-energy collision dissociation (HCD).
PRM-Based Peptide Quantification Services by Creative Proteomics
- Consultation and Experimental Design: Our team of proficient scholars shall collaborate closely with you, seeking an intimate comprehension of your research objectives and experimental prerequisites. Consequently, we shall provide adept guidance in conceiving an exemplary PRM endeavor, custom-tailored to your precise exigencies.
- Peptide Enrichment: We extend our services in peptide enrichment, elevating the sensitivity and selectivity of PRM analyzation. Utilizing a myriad of enrichment strategies, encompassing immunoaffinity purification, peptide fractionation, and phosphopeptide enrichment, we successfully amplify target peptides from intricate biological specimens.
- PRM Experiment Setup: Amongst our ranks, dwell distinguished scientist's adept in orchestrating and refining the parameters of PRM experiments. These parameters include instrument configuration, peptide fragmentation techniques, and acquisition settings, all assuring a precise and replicable outcome.
- Mass Spectrometry Analysis: Creative Proteomics prides itself on its utilization of cutting-edge mass spectrometry apparatus, predominantly high-resolution quadrupole Orbitrap systems. Such instruments amass PRM data with exceptional sensitivity and precision. Our experts execute targeted data acquisition, skillfully electing specific peptide precursor ions and meticulously monitoring their fragment ions.
- Data Processing and Analysis: Employing advanced algorithms and software, our accomplished bioinformatics specialists diligently process and scrutinize acquired PRM data. Proprietary software, adept at peak integration, precise quantification, and statistical analysis, engenders comprehensive reports of utmost accuracy.
- Quality Control and Validation: In our ardent commitment to ensuring dependable and precise outcomes, we meticulously implement quality control measures. Replicates, internal standards, and rigorous validation against reference standards or other orthogonal quantification methodologies serve as significant components of this meticulous endeavor.
Techniques and Instrumentation for PRM-Based Peptide Quantification Services
Techniques and Instrumentation for PRM-based peptide quantification analysis service include:
- Liquid Chromatography (LC): Separation of peptides based on their physicochemical properties using a variety of LC technologies such as nanoLC or UHPLC.
- Thermo Scientific Q Exactive series (Q Exactive, Q Exactive Plus, Q Exactive HF)
- Waters Xevo TQ-XS
- Agilent 6495 QQQ
- Mass Spectrometry (MS): Analysis of peptides by ionizing them and measuring their mass-to-charge ratio.
- Thermo Scientific Orbitrap Fusion Lumos Tribrid
- AB SCIEX TripleTOF 6600
- Shimadzu LCMS-8060
- Data Acquisition: Use of PRM mode on a mass spectrometer to selectively monitor and quantify predefined target peptides.
Analysis Methods of PRM-Based Peptide Quantification
Creative Proteomics employs various advanced analysis methods for PRM-Based Peptide Quantification:
- Targeted Peptide Selection: We work closely with you to identify the specific peptides of interest for quantification based on their relevance to your research objectives and biological pathways of interest.
- Transition Selection: Our experts utilize in silico tools to predict and select the optimal peptide and fragment ion transitions for PRM analysis, ensuring high sensitivity and specificity.
- PRM Data Acquisition: We configure the mass spectrometer to perform PRM data acquisition, allowing for the precise and simultaneous quantification of multiple peptides of interest.
Our Advantages of PRM-Based Peptide Quantification Services
- High Selectivity: PRM-Based Peptide Quantification enables the specific monitoring and quantification of target peptides, even in complex biological samples.
- High Sensitivity: PRM technology offers exceptional sensitivity, allowing for the detection and quantification of low-abundance peptides.
- Accuracy and Precision: PRM combines the benefits of high-resolution and accurate-mass instruments, providing accurate and precise quantification results.
- Multiplexing Capability: PRM allows for the simultaneous quantification of multiple peptides, enabling the analysis of complex biological systems.
* For research use only. Not for use in diagnostic procedures!
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