Pregled bibliografske jedinice broj: 850527
N- and O-Glycomics from Minor Amounts of Formalin- Fixed, Paraffin-Embedded Tissue Samples
N- and O-Glycomics from Minor Amounts of Formalin- Fixed, Paraffin-Embedded Tissue Samples // High-Throughput Glycomics and Glycoproteomics Methods and Protocols / Lauc, Gordan ; Wuhrer ; Manfred (ur.).
New York (NY): Springer, 2017. str. 131-145
CROSBI ID: 850527 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
N- and O-Glycomics from Minor Amounts of Formalin- Fixed, Paraffin-Embedded Tissue Samples
Autori
Hinneburg, Hannes ; Schirmeister, Falko ; Korać, Petra ; Kolarich, Daniel
Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, znanstveni
Knjiga
High-Throughput Glycomics and Glycoproteomics Methods and Protocols
Urednik/ci
Lauc, Gordan ; Wuhrer ; Manfred
Izdavač
Springer
Grad
New York (NY)
Godina
2017
Raspon stranica
131-145
ISBN
978-1-4939-6491-8
Ključne riječi
Formalin-fixed, paraffin-embedded (FFPE); Glycomics; Glycoprotein; Laser capture microdissection; Liquid chromatography; Mass spectrometry; N-glycan; O-glycan; Porous graphitized carbon
Sažetak
The availability of well-defined samples in sufficient numbers represents a major bottleneck for any biomarker related research. The utilization of preserved, archived and clinically well-described samples therefore holds a great potential to bridge this gap. This chapter describes a universal workflow for the comprehensive characterization of N- and O- glycans released from whole formalin-fixed, paraffin-embedded tissue sections, including an option for further partitioning using laser microdissection of specific tissue areas/cell populations. Glycoproteins are extracted and subsequently immobilized onto a PVDF membrane prior enzymatic release of N-glycans. Following N-glycan retrieval O-glycans are released using reductive β-elimination from the same sample spot, significantly reducing the required amount of starting material. Released and reduced glycan structures are characterized using porous graphitized carbon liquid chromatography online coupled to an electrospray ionization-ion trap mass spectrometer. This technique provides information on the relative abundances of individual glycans along with detailed structural information, including isomer differentiation and functional epitope characterization of N- and O-glycans obtained from minimal amounts of tissue down to a few thousand cells.
Izvorni jezik
Engleski
Znanstvena područja
Biologija
