Cy5-Fuc Labeled Glycan G2f

Name: Cy5-Fuc Labeled Glycan G2f
Brand: R&D Systems
SKU: GL302

R&D Systems, part of Bio-Techne | Catalog # GL302

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Product Datasheet / COA / SDS

Key Product Details

Key Benefits:

Learn more about Fluorescent Glycan Labeling and Detection

Summary for Cy5-Fuc Labeled Glycan G2f

Formula	Fuc1Gal2Man3GlcNAc4Cy5
Molecular Weight	2616.44 Da
Formulation	Supplied in 20 mM Tris, pH 8.0
Stability & Storage	Store at

Applications

Used as a ligand for lectins and to study glycan protein interaction.
Used as a substrate for various glycosidases and glycosyltransferases.
Used as a scaffold for the synthesis of various glycan epitopes such as sialyl Lewis X structures.

Key Features and Benefits:

Excitation at 649 nm and emission at 671 nm.
The fluorescent dye Cy5 is conjugated to the C6 position of the core-6 fucose.
Can be separated on 15%-17% SDS-PAGE and directly visualized as a single band under a fluorescent imager.
Linear response range for Cy5-labeled glycans can be from 10 fmol to 100 pmol, depending on the sensitivity of detection.
Store the unopened product at

For Details:

Related Reagents

Click Chemistry

Enzymes and detection reagents

Data Examples

Recombinant Sp Endo-S2 assay using Cy5-G2f (Catalog # GL302) as a substrate. Cy5-G2f (0.2 pmol per reaction) was digested by a serial dilution of Endo-S2 for 60 minutes at 37^oC. The digestions were then separated on a 17% polyacrylamide gel and imaged with TCE staining (top panel) and a fluorescent imager (lower panel). Endo-S2 digestion on Cy5-G2f resulted in two products: unlabeled product 1 and labeled product 2. Product 2 is visible with 25 ng of Endo-S2 and achieves the maximal level at 100 ng of the enzyme.

Scientific Data Examples for Cy5-Fuc Labeled Glycan G2f

	Cy5-Fuc Labeled glycan G2f This substrate can be used as ligand for lectins and to study various glycosidases and glycosyltransferases. This can also be used as a scaffold for the synthesis of various glycan epitopes such as sialyl Lewis X structures. The fluorescent dye Cy5 is conjugated to the C6 position of the core-6 fucose.

Preparation & Storage

Storage

Store the unopened product at -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Assay Procedure

Sample Assay Protocol of using Cy5-Fuc Labeled Glycan G2f/Cy5-G2f as substrate for recombinant Sp. Endo-S2 assay

Protocols are guidelines. Parameters need to be optimized by end users. Suggested input of a Cy5-labeled glycan in an assay separated on SDS-PAGE is from 0.01 pmol to 1 pmol.

Materials

Assay Buffer: 50 mM MES, pH 6.0.
Recombinant Sp. Endo-S2
Cy5-Fuc Labeled Glycan G2f (Cy5-G2f) (Catalog # GL302)
6X gel loading dye
A fluorescent imager

Assay

Dilute rSp. Endo-S2 to 10 ng/µL in Assay Buffer.
Dilute Cy5-G2f to 0.02 µM in Assay Buffer.
Mix 10 µL of the diluted rSp.Endo-S2 and 10 µL of the Cy5-G2f in a centrifuge tube.
Prepare a negative control by mixing 10 µL of the Cy5-G2f with 10 µL of Assay Buffer.
Incubate the reaction and control at 37^oC for 60 minutes.
Stop the reactions and controls by adding 5 uL of 6X gel loading dye to each tube.
Load 12.5 µL each of the above samples per well on a 15% SDS-PAGE and run at 10 volts/cm until the dye front is more than two thirds of the gel.
Image the gel with a fluorescent imager for 10 seconds.
Determine percent digestion with Alphaview or any other densitometry software.

Final Assay Conditions Per Reaction

rSp. Endo-S2: 100 ng

Cy5-G2f: 0.2 pmol

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