LudgerLiberate hydrazinolysis kit
The kit contains reagents for the release of N and O-linked glycans from glycoproteins via hydrazinolysis.
The LudgerLiberate Hydrazinolysis Kit contains reagents for the release of N and O-linked glycans from glycoprotein biopharmaceuticals. Released glycans have free reducing terminii to allow fluorescent tagging by reductive amination. The release conditions can be optimized for release of N-glycans, O-glycans or both N- and O-glycans. An alternative method for the release of O-glycans is the LudgerLiberate ORELA Kit (LL-ORELA-A2).
The kit contains reagents and materials for up to 12 glycoprotein samples analysed in parallel or two sets of 6. Up to 1 mg of glycoprotein can be deglycosylated per sample.
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Application : For release of N + O-linked glycans from glycoprotein therapeutics
Description: The kit contains reagents for the release of N and O-linked glycans from glycoprotein biopharmaceuticals. Released glycans have free reducing terminii to allow fluorescent tagging by reductive amination. The release conditions can be optimized for release of N-glycans, O-glycans or both N- and O-glycans.
Number of Samples: The kit contains reagents and materials for up to 12 glycoprotein samples analysed in parallel or two sets of 6.
Amount of Sample: Typically, up to 1 mg of glycoprotein per sample.
Suitable Samples: Biopharmaceutical glycoproteins.
Storage: Store refrigerated at 4 - 10°C in the dark. If you have limited cold storage space then store just the CEX cartridges at 4°C and the rest of the kit at room temperature. Protect from sources of heat, light, and moisture. When stored correctly, the reagents should be stable for at least 18 months from date of manufacture.
Shipping: The product should be shipped at ambient temperature. As it contains small amounts of anhydrous hydrazine the kit must be packaged and shipped as dangerous goods (DG).
For research use only. Not for human or drug use
Additional Reagents and Equipment Required
- Pure water: resistivity 18 Mμ-cm, particle free (>0.22 μm), TOC <10 ppb
- Dialysis membranes, PD10 columns, centrifugal concentrators§ or similar for removal of salts and detergents from your glycoprotein samples*
- Glove box with dry, oxygen-free inert gas (e.g. nitrogen or argon) for handling anhydrous hydrazine. As an alternative to a glove box, hydrazinolysis can be performed in a dry fume hood with inverted funnel applying a blanket of dry argon.
- Syringe (glass or PTFE) to transfer anhydrous hydrazine – e.g. 1 ml Hamilton or SGE glass syringe for liquids with teflon tipped plunger and stainless steel or PTFE needle. Do not use plastic syringes.
- Heating block, oven, or similar dry heater (a water bath cannot be used) that can be set at between 40°C and 100°C
- Centrifugal evaporator (e.g.ThermoSavant SpeedVac®, Heto or GeneVac®). If using the ThermoSavant SpeedVac® we recommend the use of the Thermo Savant RH32-13 Rotor.
This kit contains the following materials and reagents:
|LL-HYDRAZINE-01||Hydrazinolysis Release Reagent||2 x 3ml|
|LL-BICARB-01||Solid sodium bicarbonate||2 x 0.33 g|
|LL-OCTANOL-01||Octanol||2 x 0.2 ml|
|LL-ACETANHYD-01||Acetic anhydride||2 x 0.5 ml|
|LL-TFA-5PC-01||TFA solution||2 x 4 ml|
|LC-EB20-01||LudgerClean™ EB20 cartridges||2 x 6 cartridges|
|LC-EB20-WASHA-01 EB20||Wash A Solution||2 x 12 ml|
|LC-EB20-WASHB-01||EB20 Wash B Solution||2 x 12 ml|
|LC-CEX-H-01||LudgerClean™ CEX cartridges||2 x 6 cartridges|
|LL-WASTEV-01||Wash waste vials||2 x 6 vials|
|LL-REACT-01||Glass reaction vials with PTFE lined caps||2 x 6 vials|
|LL-COLLV-01||Glass collection vials with PTFE lined caps||2 x 6 vials|