Sample Preparation Kits

for Glycan Analysis

Our glyXprep sample preparation kits for glycan analysis allow effortless, straightforward, and reproducible preparation of up to hundreds of samples with minimal hands-on time.

glyXprep is structured in a modular fashion and comes in various size options to cover a wide range of applications and sample types – including N-glycan profiling of purified glycoproteins such as monoclonal antibodies, or complex glycoprotein mixtures like blood serum and plasma, and even whole-cell lysates. glyXprep is also suitable for preparing samples containing non-protein-bound free oligosaccharides as found in milk and other biofluids.

up to

Samples per Kit

under

hours

Preparation Time

under

hours

Hands-on Time

Modular Workflow

The glyXprep sample preparation workflow consists of three separate modules covering specific aspects of the sample treatment process.

These include enzymatic release of N-glycans from glycoproteins, followed by labeling of the glycans using a dedicated fluorescent dye, and lastly a post-labeling clean-up step to remove excess dye as well as residual proteins and other contaminants from the sample.

N-Glycan Release

In order to perform glycoprofiling of proteins, their glycans need to be released.

The release module of glyXprep utilizes highly specific endoglycosidase PNGase F which cleaves N-glycans from their parent protein backbone.

The glyXprep release module is compatible with all kinds of glycan profiling applications (whether CE, LC, or MS based), but is particularly optimized for use with our xCGE-LIF based high-performance glyXboxCE systems.

Glycan Labeling

Fluorescent labeling is a powerful tool to sensitively and selectively visualize glycan profiles.

The labeling module of glyXprep tags glycans with highly pure APTS fluorophore using our newly-developed, non-toxic reducing agent ReduX for maximum labeling efficiency.

APTS is exceptionally suited for capillary electrophoretic applications such as glyXera‘s high-throughput glyXboxCE systems.

Post-Labeling Clean-Up

To remove excess dye, salts, and other contaminants, a dedicated post-labeling clean-up is of importance.

The purification module of glyXprep employs highly efficient hydrophilic interaction chromatography based solid phase extraction (HILIC-SPE) to purify APTS labeled glycans.

Variants of the module exist to fine-tune the treatment for smaller sugars (e.g. milk oligosaccharides), or for larger, more complex glycan structures.

Versatility Exemplified

The glyXprep sample treatment modules are combined into ready-to-use kits to fit the requirements of a broad spectrum of glycoanalytical applications.

All glyXprep sample preparation kits contain all required buffers, reagents, and consumables to ensure best results with minimal hands-on time. Also included are detailed point-by-point instructions that guide through the sample preparation process so that even users without years-long laboratory experience can get up to speed in no time.

Our kits distinguish themselves by their ease of use while providing high sensitivity in detection and high reproducibility of results. Thanks to their well plate-based design, our kits are highly compatible with automated sample preparation systems.

Sample
Preparation Kit

N-Glycan
Release Kit

Glycan
Labeling Kit

Glycan Labeling Kit PLUS

Post-Labeling
Clean‑Up Kit

Release

Labeling

Clean-Up

Applications

Characterization of protein glycosylation

Integration into other workflows

Characterization of free oligosaccharides

Integration into other workflows

Frequently Asked Questions

What types of samples can be prepared?

Our kits have a wide range of applications, from pure proteins like monoclonal antibodies, hormones, enveloped viruses, up to complex samples like human blood plasma and serum, intact cells, or tissues.

However, for very complex samples, occasionally special sample pre-treatment is necessary. For more information, please contact our application specialists.

How much sample do I need?

The amount of required sample varies depending on the type of sample, the variant of kit used and for what analytical method the samples are prepared.

As a general rule, highly sensitive methods, like our high-performance glyXboxCE system, typically require sample amounts between 2 and 20 µg of purified glycoprotein, whereas other less sensitive methods may need an order of magnitude more sample. To get a rough estimate of the needed amount of glycoprotein sample, feel free to use the calculator tool below:

Using the labeling module for free oligosaccharides, between roughly 0.01 and 0.25 µg of oligosaccharide sample are recommended, depending on the complexity of the sample.

All our kits come with a comprehensive User Guide that details many common use cases for a variety of applications and sample types. If you need further assistance, please feel free to consult with our application specialists.

Do I need special equipment to use the kits?

Yes. All enzymes, buffers, solutions, and controls are included in the kits, but basic lab equipment like pipettes, pipette tips, reaction tubes and a temperature-controlled environment should be available at the customer.

Moreover, a well plate vacuum manifold and vacuum pump are required for the sample clean-up module. A list of compatible vacuum manifolds can be provided upon request.

How long can samples be stored after the clean-up step?

Samples are stable for several years after preparation. After the clean-up using glyXbeads, samples are highly pure with a very low conductivity. Based on internal testing we did not observe any appreciable degradation of samples when stored at -20°C over a period of multiple years. Moreover, prepared samples are very resilient against repeated freeze and thaw cycles.

Do the kits have an expiration date?

Yes. All our kits are guaranteed to have an expiration date between 6 and 12 months after delivery.

The kit component with the shortest expiration date is the endoglycosidase PNGase F of the N-glycan release module. While the enzyme is provided in lyophilized form, and therefore suitable for long-term storage, once reconstituted, it remains stable for about 3 months. Multiple freeze-thaw cycles are not recommended to preserve optimal enzyme activity.

Show More

Do the kits contain any highly toxic components?

No. Special care has been taken to enable kit use in a low safety level environment. No hazardous or toxic ingredients are contained in the kits.

Does my sample have to be in a special buffer or under special conditions?

Our kits are compatible with a wide range of sample buffers at pH around 7.2, like PBS or Tris. If you plan to use a buffer system different from the recommended ones, we advise you to evaluate kit performance in advance, or consult with our application experts.

For very low or high sample pH values or salt concentrations, it is possible to perform a buffer exchange via diafiltration against PBS.

Which analysis methods are compatible with the kits?

Our kits are compatible with a wide range of methods. While our kits are optimized for use with CE or CGE based separation methods, they can also be used with HPLC or UPLC applications, for example, using HILIC or C18 columns.

If you need more information about compatible CE and LC applications and possible buffer systems, please contact our application specialists.

Can I also analyze whole cell N-glycosylation using the kits?

Yes, our kits can handle complex samples just fine. For best results, simply ensure an adequate cell disruption by using a RIPA buffer and/or mechanical shear stress with a syringe.

Still have more questions?

If you are unsure which kit is the right one for your project, or if you are interested in a tailor-made solution for your specific application, please feel free to get in touch with our experts and we are happy to help you with any questions you have.

Get in Touch