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Proteomics refers to the totality of all proteins present in a living organism, tissue, cell or cell compartment at a certain point in time under defined conditions. The proteome is subject to constant change, as proteins are constantly being synthesised or degraded. Proteomics analysis is therefore mainly concerned with sample preparation and the separation of proteins, enzymes, peptides and amino acids. It also includes RNA and DNA analysis in some cases.
Sample preparation primarily includes protein extraction, peptide purification and phosphopeptide enrichment.
A range of chromatographic separation methods are available for analysing proteins and peptides. These include reversed-phase chromatography, hydrophilic and hydrophobic interaction chromatography (HILIC and HIC), ion exchange chromatography (IEX), size exclusion chromatography (SEC) and affinity chromatography (AFC).
Below you will find further information and catalogue material with application examples from various manufacturers for the analysis of proteins, peptides and amino acids as well as DNA, RNA and other related substances.
Below you will find more detailed information with application examples for analysing specific proteins, peptides, DNA, RNA, antibodies, amino acids, etc.
A large number of proteins and peptides play an important role in the analysis of proteomics. Mass spectrometry (MS) is often used to detect these analytes. There are two different approaches for the MS analysis of proteomics: top-down and bottom-up.
Top-down is used to analyse and fragment intact proteins by ESI-MS (Electron Spray Ionisation) in the gas phase. The large number of different charge variants of the proteins can lead to complicated spectra. Liquid chromatography can help to separate complex samples before they are detected by ESI-MS. The top-down approach can be used to recognise degradation products and sequence variants, which help to solve interference problems and determine post-translational modifications.
Bottom-up describes the approach in which the proteins are first enzymatically cleaved (also called digestion, often tryptic digest). The resulting peptides can then be analysed by mass spectrometry. Individual proteins or a mixture of proteins can be cleaved. This enzymatic cleavage yields thousands to hundreds of thousands of peptides, which is why a two-dimensional approach can be helpful. The original proteins can then be identified by comparison with theoretical peptide masses or a database.
Sample preparation - Tryptic Digest:
Sample preparation - Desalting and enrichment:
Sample preparation - Phosphopeptide enrichment:
Sample preparation - Desalting of phosphopeptides:
Sample preparation - removal of phosphopeptides:
Sample preparation - purification and enrichment of small samples:
Sample preparation - peptide fractionation:
Sample preparation - antibody purification:
Proteomics - Reversed Phase:
Proteomics - HILIC:
Proteomics - SEC:
Proteomics - IEX:
Proteins and peptides - Reversed phase:
Proteins - SEC:
Proteins, peptides and antibodies - SEC:
Antibodies - IEX:
Antibodies - Affinity chromatography:
Proteomics - IEX:
Proteomics - Reversed Phase:
Proteomics - HIC:
mRNA - Affinity chromatography:
Adeno-associated Viruses - IEX:
Proteins and peptides - SEC:
Proteins and Peptides - Reversed Phase:
Proteins and peptides - HILIC:
Proteins and peptides - IEX:
Proteins and peptides - mixed mode:
Amino acids - IEX:
Amino acids - Reversed phase:
Amino acids - HILIC:
Nucleotides - Mixed-Mode:
Nucleotide - Reversed Phase:
Oligonucleotides - HILIC:
DNA/RNA - IEX:
DNA/RNA - SEC:
Peptides - Reversed Phase:
Proteins - Reversed Phase:
Proteins - HIC:
Proteins - Charge variants with IEX:
Antibodies - affinity chromatography:
Antibodies - SEC:
Antibody - Reversed Phase:
Antibodies - IEX:
Glycans - HILIC:
Glycans - Mixed-Mode:
Peptide Mapping - Reversed Phase:
Nucleic acids and oligonucleotides - reversed phase:
Oligonucleotides - IEX:
Proteins and peptides - metal affinity:
Sample preparation - Digestion:
Proteins - IC:
Proteins and peptides - SEC:
Amino acids - IEX:
Peptides and small proteins - IEX:
Peptides - HIC:
Proteins - IEX:
Proteins - HIC:
Proteins - Reversed Phase:
Nucleosides - IEX:
Nucleotides - IEX:
Peptides, oligonucleotides, nucleobases - HILIC:
Oligonucleotides - IEX:
Oligonucleotides and DNA/RNA - HIC:
Oligonucleotides and DNA/RNA - Reversed Phase:
DNA/RNA - IEX:
Antibody affinity chromatography:
Peptides - UHPLC - Reversed Phase:
Peptides - Reversed Phase:
Peptides/Proteins - IEX:
Proteins - Reversed Phase:
Proteins - UHPLC - HILIC:
Proteins - UHPLC - HIC:
Proteins - HIC:
Proteins - UHPLC - IEX:
Proteins - UHPLC - Reversed Phase:
Proteins - UHPLC - SEC:
Proteins - SEC:
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