Protein Quantification Service-Wuhan Biobank Co., Ltd.

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Protein Quantification Service

Product Description

Case Analysis

Results Display

Sample Submission Guidelines

Q&A

a.TMT™ Technology

TMT™ (Tandem Mass Tag™) technology is an in vitro peptide labeling technology developed by Thermo Scientific, which is designed for the identification and quantification of proteins in different samples by tandem mass spectrometry (MS). This technique uses multiple isotope labels to label the amino groups of peptides, and after LC-MS/MS analysis, the relative amounts of proteins in 2-16 groups of different samples can be compared simultaneously.

TMT ™ Technical schematic diagram

Instruments used:Orbitrap Eclipse, Orbitrap Exploris 480,Q Exactive Plus.

Technical advantages: Simultaneous analysis of protein differences in 2-16 samples of different conditions. Suitable for a wide range of sample types: cytoplasmic proteins, membrane proteins, nuclear proteins, extracellular proteins, etc. Compared with protein quantification by gel separation, TMT mass spectrometry can detect more types of protein molecules, such as low-abundance proteins, strongly acidic (base) proteins, <10KD or >100KD protein molecules.

Application areas: disease marker screening, mechanism of action research, plant resistance research, drug action target research, special functional protein screening.

Sample requirements

Sample Type	Sample requirements (per group of samples)
Protein extract	Concentration >1μg/μL, total protein >300μg
Cell sample	Cell volume >10^7
Tissue samples	Animal, microbial tissue wet weight >10mg; plant fresh tissue >100mg
Body fluid sample	Blood volume>500μL

　Lead time：

Progress	Time period
Sample collection/Quality control	1-2 working days
Sample processing	5-6 working days
Mass spectrometry test	3-4 working days
Data analysis	2-3 working days
Total	No more than 20 working days

Note: For more than 10 samples, 1 working days will be added for each additional sample. If there is an urgent need, please contact us for negotiation.

b.Label-free

Label-free quantification is the mass spectrometric analysis of enzymatic protein peptides by liquid chromatography-mass spectrometry (LC-MS).

Label-free quantitative proteomics experimental process

Instruments used: THERMO: Orbitrap Eclipse, Orbitrap Exploris 480,Q Exactive Plus.

Technical advantages: no labeling of proteins, low cost and low volume of samples required.

Application areas: disease marker screening, mechanism of action research, plant resistance research, drug target research, special function protein screening.

c.SWATH/DIA

SWATH/DIA is a new panoramic scanning mode for mass spectrometry. Compared with the traditional shot-gun technique, SWATH/DIA acquisition mode is able to scan all precusor peptide in the scan interval at ultra-high speed and perform secondary fragmentation, thus obtaining complete peptide information. With SWATH /DIA acquisition mode, complete quantitative and qualitative results can be obtained in a single experiment, without method optimization. SWATH/DIA mode overcomes the problem of shot-gun scan randomness, resulting in better data reproducibility. SWATH/DIA is particularly suitable for enrichment of purified samples, e.g. SWATH/DIA quantification after IP to detect proteins that interact with the target protein.

SWATH technical schematic diagram

Instruments used: THERMO: Orbitrap Eclipse, Orbitrap Exploris 480, Q Exactive plus; AB SCIEX: TripleTOF 5600 plus

Technical advantages

High sensitivity, which has the data acquisition mode of MRM, combined with a high-resolution mass spectrometry system with comparable sensitivity to MRM.

Quantitative correlation between duplicate samples can reach above 0.99.

Quantitative accuracy almost comparable to MRM technique.

the quantification range can span 4 orders of magnitude

The quantitative results are very good.

Application areas: mechanism and regulatory mechanism research, screening of disease markers, screening of drug use and prognostic markers, molecular typing of diseases.

Sample requirements：

Sample Type	Sample requirements (per group of samples)
Protein extract	Concentration >1μg/μL, total protein >300μg
Cell sample	Cell volume >107
Tissue samples	Animal, microbial tissue wet weight >10mg; plant fresh tissue >100mg
Body fluid sample	Blood volume>500μL

Lead time:

Progress	Time period
Sample collection/Quality control	1-2 working day
Sample processing	5-6 working days
Mass spectrometry test	3-4 working days
Data analysis	2-3 working days
Total	No more than 20 working days