Orela O-linked Glycan Release Kit
The Ludger Liberate Orela Kit has been developed for the release of O-linked glycans from glycoproteins. Released glycans have free reducing terminii to allow fluorescent tagging by reductive amination.
Part Number: LL-ORELA-A2
The kit includes reagents and materials for up to 12 glycoprotein samples analysed in parallel or two sets of 6 samples, typically up to 1 mg of glycoprotein per sample.
– Designed for easy, routine analysis of O-glycosylation by QC labs.
– Straightforward and simple method.
– Released glycans have free reducing terminii suitable for fluorescent labelling prior to analysis by HPLC and MS.
– Fluorescent labelling allows relative quantitation of glycan species by HPLC.
– Faster and lower cost than hydrazinolysis.
– No special handling techniques.
The Orela Reaction
The Orela reagent reacts at the link between the glycan and peptide backbone to liberate glycans as the glycosylamine. The acid-labile glycosylamine derivative is hydrolysed to produce the free glycan.
Orela Kit Components
|LL-ORELAREAGENT-0||Orela Release Reagent||2 x 3ml|
|LL-ACETIC-50P-01||Acetic Acid Solution||2 x 3ml|
|LC-CEX-H-01||LudgerClean CEX-H cartridges||2 x 6 cartridges|
|LL-COLLV-01||Collection vials with PTFE lined caps||2 x 6 vials|
|LL-REACT-01||Reaction vials with PTFE lined caps||2 x 6 vials|
Outline of the Orela Protocol
Prepare the glycoconjugate
Prepare the glycoprotein or glycopeptide samples by removing contaminants such as salts, detergents and dyes that could interfere with the labeling procedure.
Dry the glycans
Place the samples in reaction vials and dry down.
Add release reagent to glycoconjugates
Add Orela release reagent to each sample.
Incubate the samples to allow the release reaction to progress.
Add acetic acid solution and incubate at 4C to convert glycosylamines into non-reduced glycans
Remove the peptide or protein using a cation exchange column
Store or Analyse released glycans
The released glycans are now ready for analysis