Label free experiments

Label free experiments

Non standard quantification (label free)

Label free quantification: It is a non labeled differential proteomics technique that does not require the use of isotopic labels to label peptide segments. It directly quantifies peptides or proteins based on the signal intensity of mass spectrometry peaks. Label free analysis of enzymatically hydrolyzed peptide segments using HPLC fractionation and liquid chromatography-mass spectrometry There are currently two main non-standard quantitative techniques based on mass spectrometry: one is protein quantification based on information such as peptide peak intensity and peak area related to primary mass spectrometry, and the other is quantification based on the total number of secondary mass spectrometry (spectral counts) of the corresponding peptide segments of the protein.

Experimental Procedure

1. Protein pretreatment, including removal of nucleic acid, protein purification and quantification, SDS polyacrylamide gel electrophoresis;

2. Enzymatic cleavage of protein gel;

3. Peptide purification; .

4. Mass spectrometry analysis of peptide mixtures, including mass spectrometry identification and quantitative information comparison;

5. Bioinformatics analysis

Results Display

Experimental cycle and delivery standards

1. Experimental period: 4-6 weeks

2. Delivery standards: protein quantification results, identification results, mass spectrometry raw data, bioinformatics analysis results (G0 analysis) KEGG analysis, etc

Information to be confirmed

1. Species of the sample

2. Type of sample (tissue or cell or blood)

3. Sample quantity and grouping

4. Analysis requirements

References

[1]HuS, Qiu N, Liu Y , et al. Identification and comparative proteomic study of quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometryanalysis[J]. Poultry Science, 2016:pew033.

[2] Bouvet M , Turkieh A，Acosta-Martin A E , et al. Proteomic Profiling of Macrophages by 2D Electrophoresis[J]. Journal

of Visualized Experiments, 2014(93).

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