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BOC Sciences has a first-class experimental platform, mature mass spectrometry techniques and an experienced team of experts, and can provide comprehensive C-terminal sequencing services for various proteins, peptides, antibodies, and amino acid samples. In addition, we also provide N-terminal sequencing services.
Protein is a chain-like polymer compound formed by the dehydration condensation of amino acids, including two ends of the amino terminal (N-terminal) and the carboxyl-terminal (C-terminal). The C-terminus is an important structural and functional site of proteins and peptides, and plays a decisive role in the biological functions of proteins. The analysis of the type and sequence of C-terminal amino acids of proteins and peptides helps to reveal the structure and function of proteins. Although the N-terminal sequencing technology based on Edman degradation is becoming more and more mature, there are still many problems that still need to be solved by C-terminal sequence analysis. C-terminal sequencing is especially suitable for quality control in large-scale production, analysis of N-terminal blocking proteins and peptides, and design of DNA probes.
We use mass spectrometry to perform amino acid C-terminal sequencing services on samples provided by customers within 4-5 weeks. After enzymatic digestion, desalting and separation of protein or peptide samples, we use advanced mass spectrometers to complete mass spectrometry analysis. Obtain the m/z spectrum of the fragment ion of the peptide in the mass spectrometer, and analyze the spectrum to obtain the C-terminal amino acid sequence of the sample. The number of amino acids determined depends on the enzyme cleavage site. After the sequencing is completed, we deliver a detailed project report including information such as the C-terminal amino acid sequence, mass spectrum, relevant mass spectrometry parameters, raw data, and basic description of the experimental method.
The sample state provided by the customer can be dry powder or solution. The total amount of the sample should be greater than 300ug and the purity greater than 90%. The salt content requirements of the sample are the same as those for N-terminal sequencing, and the volatile inorganic salt content in the sample should be less than 20mM, and the non-volatile inorganic salt content should be less than 5mM.
Carboxypeptidase is an exonuclease that releases amino acid residues one by one from the C-terminus of proteins. We use carboxypeptidase to hydrolyze protein or peptide samples, and use chromatography and mass spectrometry to detect the sequence of amino acids released by carboxypeptidase one by one, and finally obtain the C-terminal amino acid sequence information. Chromatography allows direct detection of released amino acids. Mass spectrometry can determine the type of amino acid released by measuring the difference between the original molecular weight of a protein or peptide and the molecular weight after releasing an amino acid, and so on.
We obtained the C-terminal amino acid by chemical reagent cleavage method, and then combined with liquid chromatography or mass spectrometry to identify the amino acid species. After the protein or peptide reacts with chemical reagents, specific cleavage can occur.
