Fetuin Glycoprotein Standard

The fetuin glycoprotein standard was developed for use during glycan release and labeling(particularly for DMB sialic acid analysis). Fetuin exists in a variety of glycoforms containing bi-, tri-, and tetra-antennary oligosaccharides with variable sialylation, as well as O-linked glycans.

Amount of Glycoprotein Supplied
GCP-FET-05          500 µg
GCP-FET-250U     250 µg
GCP-FET-50Ux4   50 µg per vial – 4 vials

Source Fetal calf serum

Form Dry. Lyophilised powder.

Molecular Weight 36 kDa (protein weight only)

Amount of Protein
GCP-FET-05          300 µg
GCP-FET-250U     150 µg
GCP-FET-50Ux4   32 µg per vial – 4 vials
In comparison to BSA standard, determined by BCA assay. Value rounded to nearest μg.

Analysis
Fetuin glycans were released from the Fetuin Glycoprotein Standard (Cat# GCP-FET-05) using PNGase-F.

Following release the glycans were labeled using 2-Aminobenzamide (2-AB) using the LudgerTag 2-AB Glycan Labeling Kit (Cat# LT-KAB-A2).

Figure 1: Waters BEH column HPLC profile of 2-AB labeled Fetuin N-glycans, released by PNGase F from GCP-FET-05 batch B1B2-04.

Figure 1 shows a LudgerSep N2 HPLC profile of bovine fetuin N-glycans. To thoroughly investigate the N-glycans we first separate them based on charge on a LudgerSep C3 column (Figure 2) and then run each fraction on a LudgerSep N2 column.

Figure 2: LudgerSep-C3 profile and subsequent LudgerSepN2 analysis of bovine fetuin PNGaseF released N-glycans from a similar batch of GCP-FET.
2-AB labeled glycans were separated on the LudgerSepC3 column and these fractions were then separated on the LudgerSepN2 column. This figure demonstrates the complexity of N-glycans present in the sample. A combination of LudgerSepC3/N2 and exoglycosidase digestion is required to identify the glycans and their relative abundance. N- neutral glycans, 1S – monosialylated glycans, 2S – disialylated glycans, 3S – trisialylated glycans & 4S – tetrasialylated glycans. For more detailed information on glycan identification, please see the product documentation.

Storage Refrigerate (-20°C) both before and after dissolving. This product is stable for at least 5 years as supplied.

Shipping The product is shipped at ambient temperature.

Handling Once dissolved avoid repeated thawing and refreezing, storage over 3 hours at room temperature or above, exposure to light and long term exposure to acid as these will cause glycan desialylation.

Safety This product is non-hazardous and has been purified from natural sources certified to be free of all hazardous material including pathogenic biological agents.