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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.

View product documentation as PDF

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

Part #ItemQuantity
LL-ORELAREAGENT-0Orela Release Reagent2 x 3ml
LL-ACETIC-50P-01Acetic Acid Solution2 x 3ml
LC-CEX-H-01LudgerClean CEX-H cartridges 2 x 6 cartridges
LL-COLLV-01 Collection vials with PTFE lined caps2 x 6 vials
LL-REACT-01 Reaction vials with PTFE lined caps2 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

Post-release cleanup
Remove the peptide or protein using a cation exchange column

Store or Analyse released glycans
The released glycans are now ready for analysis

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