Metabolomics Creative Proteomics

Creative Proteomics Metabolomics

Keratan Sulfate Analysis Service

Keratan Sulfate Analysis Service

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Creative Proteomics, as the leader of bio-tech industry, devote to forge high quality targeted metabolomics service with advanced apparatus and professional research team. We provide high throughput, high precision and short time-consuming Keratan Sulfate Analysis Service, which could be meet all the need of our customers, to assist your scientific research.

Overview

Glycosaminoglycans (GAGs), also known as mucopolysaccharides, are negatively charged polysaccharides consisting of repeating disaccharide units. They are present in every mammalian tissue and encompass a wide range of functions determined by their molecular structure. Historically, the functions of GAG have been considered limited to cellular hydration and structural scaffolds. However, recent evidence suggests that GAG also plays a key role in cellular signaling, regulating a wide range of biochemical processes. The four main groups of GAG are classified based on their core disaccharide units, including heparin/heparan sulfate, chondroitin sulfate, dermatan sulfate, hyaluronic acid, and keratan sulfate. Among them, keratan sulfate (KS) was the only GAG type with galactose as the structural component.

Major repeating unit of glycosaminoglycans. The color coding of the constituent monosaccharides was in accordance with the SNFG term. The abbreviations are as follows: Glc represents glucose, Ido represents galactose, Gal represents galactose, N represents amine, S represents sulfate, A represents acid, and NAc represents N- acetyl.Fig 1. Major repeating unit of glycosaminoglycans. The color coding of the constituent monosaccharides was in accordance with the SNFG term. The abbreviations are as follows: Glc represents glucose, Ido represents galactose, Gal represents galactose, N represents amine, S represents sulfate, A represents acid, and NAc represents N- acetyl.

Based on tissue specificity, most importantly based on chemical structure, three classes of KS are defined, which vary depending on the degree of sulfation, the sulfation pattern, and the type of linked oligosaccharide that binds keratan sulfate to the core protein. Form I KS is an N-chain KS chain with variable degrees of sulfation (non-sulfate, mono-sulfate and di-sulfate) and is mainly present in the cornea. Type II KS is an almost completely sulfated KS chain that connects to the core protein O-type through GalNAc, and is mainly present in cartilage tissue. KS Form III is also highly sulfated, but is attached to its core protein by a 2-O-mannose bond and is the most abundant in the central nervous system.

Advantages

  • Short time-consuming
  • High sensitivity and few detection restrictions
  • High precision and good repeatability
  • High throughput
  • Customized service

Service workflow

  • Extract and purifying glycosphingolipid from cell membrane or serum
  • Release that glycosphingolipid glycan with the glucosidase
  • The total methylation of glycan
  • UPLC-HILIC-FLD analysis based on 2-AB labeling
  • Analysis of fully methylated glycans by time-of-flight mass spectrometry

Workflow for the discovery-based proteomics.Fig 2. Workflow for the discovery-based proteomics.

Sample requirement

  • Serum/plasma: 500 μl/sample
  • Repeated freezing and thawing of samples must be avoided. The serum sample should be precipitated in the collection tube for 30 minutes at room temperature, then transported to the centrifuge tube and centrifuged at 8000 rpm for 5 minutes. After centrifugation, the supernatant was equally divided into a freezing tube of 500 uL / sample.

  • Protein: 100 µg
  • Anticoagulated blood (EDTA): 1 mL
  • Anticoagulants and preservatives must be added immediately after collection and then frozen at -80 °C.

  • Urine: 1 ml/sample
  • Urine samples should be equally divided into centrifuge tubes with 1 mL per tube, each tube is added with 1/100 (w/v) sodium azide and stored at -80 °C.

  • Animal tissues: 200 mg/sample
  • Samples should be frozen in liquid nitrogen immediately and then transported to -80 °C for storage after collected.

  • Cells: ≥ 1 × 107/sample
  • Cytoactive should be terminated immediately to maintaining cell integrity.

  • Feces: 500 mg/sample

In general, to assure enough sample to fulfill the whole project, the volume of the single sample need to be offered as much as possible. The remaining samples will be stored for one year free of charge and returned at any time if necessary. All samples need to be stored and transported at -80°C and try to avoid using surfactants (SDS, Triton-X) and inorganic salts.

Clinical samples are repeated in no less than 30 cases in a single group.

Animal samples are repeated in no less than 9 cases in a single group.

Report delivery

  • Experimental procedure
  • Parameters of HPLC and MS
  • Raw data, chromatograms and mass spectra
  • Metabolites quantification data
  • Custom analysis report

Service cycle

  • Sample testing: 5-10 working days
  • Data analysis: 5-10 working days

Creative Proteomics metabolism analysis platform is committed to the all-around, reliable and accurate analysis service for a variety of target substances, which is suitable for life-science research, drug exploration, biological determination and other fields. We sincerely hope to cooperate with you to assistant your scientific research.

References

  1. W. Wang, L. Shi, Y. Qin, et al. Research and Application of Chondroitin Sulfate/Dermatan Sulfate-Degrading Enzymes. Front Cell Dev Biol, 2020, 8:560442.
  2. Z. Wang, K. Arnold, Y. Xu, et al. Quantitative analysis of heparan sulfate using isotopically labeled calibrants. Commun Biol, 2020, 3(1):425.
For Research Use Only. Not for use in diagnostic procedures.

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