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DIA Quantitative Proteomics

Almost 100% ion utilization, increased detection depth and sensitivity.
Fewer missing values, enhanced detection stability and accuracy.
Lower sample requirements, broaded detection scope.

Technology Introduction

DIA quantitative proteomics technology utilizes the unique dual TIMS design of the timsTOF series mass spectrometer and the diaPASEF® acquisition mode to achieve qualitative and quantitative protein analysis. The diaPASEF acquisition mode combines the data-independent acquisition (DIA) mode with PASEF (Parallel Accumulation–Serial Fragmentation) technology. In the diaPASEF acquisition mode, the mass spectrum scan range is divided into multiple windows based on the mass-to-charge ratio (m/z) information of all precursor ions in the MS1 spectra. Subsequently, all precursor ions in each window are sequentially fragmented in the MS2 spectra, recording their retention time, mass-to-charge ratio, ion intensity, and ion mobility. Finally, protein qualitative and quantitative analysis is conducted based on this information.

DIA proteomics combines the advantages of 4D proteomics and DIA technology while overcoming some of their inherent limitations. Based on the Bruker timsTOF HT platform, DIA proteomics enhances ion utilization and identification accuracy, significantly improves the coverage, sensitivity, and throughput of protein detection technology with less sample input and faster scanning speed.

Applications of DIA Quantitative Proteomics

Medical Research
Disease Mechanisms, Molecular Diagnosis, Biomarker Development, Drug Targets.
Animal Research
Reproductive Development, Disease Mechanisms, Nutrient Metabolism, Animal Toxicology.
Plant Research
Reproductive Development, Abiotic Stress Response, Disease Resistance, Crop Improvement.
Microbiology
Pathogenic Mechanisms, Drug Resistance, Stress-Related Proteins Screening, Environmental Impact Mechanisms.

Project Experience

dia

Average number of proteins identified
Project Workflow
1
Sample Shipment
2
Protein Extraction
3
Trypsin Digestion
4
Data Acquisition
5
Database Search
6
Data Analysis

Sample Requirements

Sample Type Samples Recommended Sample Size Minimum
Sample Size
Human/Animal Tissue Normal tissues (heart, liver, spleen, lungs, intestines, kidneys, etc.) 50mg 5mg
Fatty tissue 200mg 100mg
Brain tissue 50mg 5mg
Bone 1g 200mg
Hair 500mg 200mg
Skin 200mg 100mg
Plant Tissue Young tissue (young leaf, seedling, petal, etc.) 200mg 100mg
Mature tissue (root, stem, fruit, pericarp, etc.) 1g 500mg
Pollen 40mg 15mg
Liquid Samples Serum/Plasma (without removing high abundance proteins) 20μL 5μL
Serum/Plasma (remove high abundance proteins) 200μL 100μL
Joint fluid, Lymph fluid 200μL 100μL
Aqueous humor, Vitreous body 300μL 200μL
Cerebrospinal fluid 200μL 100μL
Ascites, Follicular fluid 100μL 50μL
Alveolar lavage fluid (BALF) 1ml 500μL
Amniotic fluid 1ml 500μL
Milk 20μL 5μL
Urine 10mL 5mL
Saliva (mammals) 1ml 500μL
Fermentation broth, Bacterial solution 10ml 5ml
Cellular supernatant 25mL 10ml
Exosome (sediment) 25μl 15μL
Microorganisms Bacteria 200mg 100mg
Fungi 300mg 150mg
Cells Primary Cells 3×10^6 1×10^6
Transmissible cells 2×10^6 1×10^6
Sperm, Platelets 2×10^7 1×10^7
Protein Protein 100μg 30μg
Biological duplicates: A minimum of 3 replicates is required; 3-6 replicates for animal samples; 6-10 for clinical samples.

 

WHAT'S NEXT IN OMICS: THE METABOLOME

Please submit a detailed description of your project. We will provide you with a customized project plan metabolomics services to meet your research requests. You can also send emails directly to support-global@metwarebio.com for inquiries.
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