Thermo Scientific Pierce Glycoprotein Isolation Kits, Concanavilin A (ConA) and Wheat Germ Agglutinin (WGA) allow isolation of glycoproteins from complex mixtures, including serum, tissue and cultured cell lysates.
These two kits use lectins concanavilin A (ConA) and wheat germ agglutinin (WGA), respectively, as ligands to bind specific classes of glycoproteins from complex protein samples. ConA lectin recognizes alpha-linked mannose and terminal glucose residues, while WGA lectin selectively binds to N-Acetyl glucosamine (GlcNAc) groups and to sialic acid. The kits include ConA or WGA beaded agarose resin, optimized binding/wash and elution buffers, and easy-to-use microcentrifuge spin columns. The kits have been used to successfully isolate glycoproteins from serum as well as HeLa and CHO cell lysates in approximately 50 minutes.
Highlights:
- High recovery – Equivalent or greater glycoprotein recovery versus competitor kits and lectin resins
- Fast – Glycoprotein purification in less than an hour
- Versatile – Isolate glycoproteins from various sample types, e.g., human serum and cell lysate.
- Robust – Lectin does not leach from resin when processing sample
- Convenient – Complete kit offering lectin resins and spin columns with all necessary reagents
- Compatible with Bradford-based protein assays – dialysis or protein precipitation of recovered glycoproteins is not required prior to protein assay
Product Details:
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| Thermo Scientific Pierce Glycoprotein Isolation Kit Protocol |
The Concanavalin A lectin recognizes α-linked mannose and terminal glucose residues, while the Wheat Germ Agglutinin lectin selectively binds to N-Acetyl glucosamine (GlcNAc) groups and to sialic acid. The immobilzed ConA and WGA lectins used in these Glycoprotein Isolation Ktis allows selective recovery and downstream analysis of glycosylated proteins which are known to have important biological function in immune regulation, inflammation, cell-to-cell adhesion, and cell signaling.
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Glycoprotein isolation from human serum and cell lysate – performance comparison of kits using WGA resin. CHO lysate and human serum samples were processed with the Thermo Scientific Pierce Glycoprotein Isolation Kit, WGA and with other commercially available WGA resins. An equivalent amount of total protein was applied to each resin. Eluted glycoprotein fractions were normalized by volume and resolved on 8-16% polyacrylamide gels. (a) Eluted glycoprotein fraction from applied CHO lysate (b) Eluted glycoprotein fraction from applied human serum. |
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Glycoprotein isolation from human serum and cell lysate – performance comparison of kits using ConA resin. CHO lysate and human serum samples were processed with the Thermo Scientific Pierce Glycoprotein Isolation Kit, ConA and with other commercially available ConA resins. An equivalent amount of total protein was applied to each resin. Eluted glycoprotein fractions were compared with ConA Resin boiled in SDS-PAGE Buffer to release lectins. All fractions were normalized by volume and resolved on 8-16% polyacrylamide gels. Gels were then silver stained. (a) Eluted glycoprotein fractions from applied CHO lysate (b) Eluted glycoprotein fractions from applied human serum. Arrows identify protein bands that result from ConA leaching from the resin during the elution process.
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Related Resources:
A review of sample preparation for mass spectrometry
An overview of glycosylation and methods to analyze glycoproteins and glycopeptides
Related Products:
Phosphoprotein Enrichment Kit
Phosphopeptide Isolation Kit
Ubiquitin Enrichment Kit
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