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Promega Corporation

PTM-Works

ptm-works

PTM-Works is a range of post-translational modification (PTM) identification and quantitation services. These services include screening a sample for the presence or absence of a particular PTM on a target protein, global analysis of a sample to identify any proteins harboring a specific modification, and comparing the abundance of modified peptides between samples.

PTM PROFILING

This service is used to detect post-translational modifications (PTMs) on proteins. The service can be applied to purified proteins, recombinant proteins or proteins in complex mixtures. Samples should be submitted as gel bands. The range of modifications is limited to those readily identifiable in a database search, such as methylation of lysine or arginine, ubiquitination of lysine, deamidation of glutamine or asparagine, glycation of lysine, phosphorylation of serine, threonine or tyrosine (without enrichment), or acetylation of lysine.

In more complex samples, fractionation can be employed, the extent of the fractionation correlates with the number of proteins and their PTMs that are detected. The greater the number of fractions, the greater the number of PTMs identified.

Following trypsin digestion the peptide mixture is analyzed by LC/MS/MS. The peptides are fragmented in the mass spectrometer to yield diagnostic patterns that can be matched to protein sequence databases via computer algorithms.

A 10 fraction analysis is suitable for the analysis of low complexity samples e.g. HPLC fractions, semi-purified extracts. A 20 fraction analysis is for medium complexity samples e.g. subcellular fractions, pooled HPLC fractions. A 40 fraction analysis is suitable for complex samples e.g. whole cell lysates, whole tissue extracts.

SERVICE ID SERVICE DESCRIPTION LIST PRICE PER SAMPLE
MSB-9 PTM Profiling Protein $310

Sample types: 2D gel spots, SDS-PAGE bands, recombinant proteins, purified proteins, cell lysates, tissue homogenates, biofluids, FFPE extracts
Data turnaround time: Two weeks or less
Report format: PDF report

PTM Profiling Plus

This service uses multiple enzymes to maximize the sequence coverage obtained for your protein of interest. This approach increases the likelihood of detecting PTMs on your protein including those on none tryptic peptides. The range of modifications is limited to those readily identifiable in a database search, such as methylation of lysine or arginine, ubiquitination of lysine, deamidation of glutamine or asparagine, glycation of lysine, phosphorylation of serine, threonine or tyrosine (without enrichment), or acetylation of lysine.

Samples are submitted as excised gel bands. If you prefer can we run the gel for you. The gel piece is processed using trypsin/chymotrypsin and elastase, the peptide mixture is analyzed by LC/MS/MS. Data are processed using the database search tool Mascot configured to identify your PTM of interest. Mascot search results are visualized using Scaffold from Proteome Software.

SERVICE ID SERVICE DESCRIPTION LIST PRICE PER SAMPLE
MSB-10 PTM Profiling Protein Plus $900

Phospho-screen

This targeted workflow uses peptide level TiO2 enrichment to select for phosphorylated species. The phosphorylated peptides may be profiled in a single sample or monitored for differential analysis across multiple samples.

SERVICE ID SERVICE DESCRIPTION LIST PRICE PER SAMPLE
MSB-11 Phospho-screen $900

Sample types: Lysates, cells, tissues, biofluids, FFPE protein extracts, SILAC
Data turnaround time: Two weeks or less
Report format: PDF report

Phospho-screen Plus

This workflow can be used to study phosphorylated species from complex samples such as cell lysates, tissues and biofluids. Samples are processed by trypsin digestion and phosphorylated peptides are enriched using titanium dioxide microparticles before further fractionation using strong cation exchange chromatography. This service uses 20 hours of instrument time versus the 2 hours used for Phospho-screen. The increased instrument time is reflected in the number of phospho protein/peptide identifications.

SERVICE ID SERVICE DESCRIPTION LIST PRICE PER SAMPLE
MSB-12 Phospho-screen plus $2,400

Glyco-screen

This screening service can be used to determine presence or absence of N-glycosylation sites on an antibody, recombinant or highly purified protein. Modified asparagine (Asn) residues are converted to the deamidated aspartic acid (Asp) following treatment with PNGase F, this is a simple modification to pick up in a database search. Analysis without treatment accounts for any artifactual deamidation.

SERVICE ID SERVICE DESCRIPTION LIST PRICE PER SAMPLE
MSB-13 Glyco-screen $1,000

Sample types: Recombinant proteins, purified proteins (in solution or gel)
Data turnaround time: Two weeks or less
Report format: PDF report

Glyco-screen Plus

This screening service can be used qualitatively or quantitatively to determine the presence of N-glycosylation sites in complex mixtures such as cell lysates or biofluids e.g plasma and CSF. Samples are digested with trypsin and glycosylated peptides are enriched from the complex background using hydrazide chemistry. Glycosylated peptides are captured and retained on a solid support while none glycosylated proteins are washed away. PNGase F treatment is used post-capture to release N-linked glycans from the solid support. Released peptides are analyzed by LC/MS/MS.

SERVICE ID SERVICE DESCRIPTION LIST PRICE PER SAMPLE
MSB-14 Glyco-screen Plus $5,775

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Scientists at Your Service

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