Cy5-Fuc Labeled Glycan G2f

Catalog # Availability Size / Price Qty
GL302-100
Cy5-Fuc Labeled glycan G2f
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Cy5-Fuc Labeled Glycan G2f Summary

 

Key Benefits

Learn more about Fluorescent Glycan Labeling and Detection

Cy5 Labeled Antibody Glycan G2

Formula

Fuc1Gal2Man3GlcNAc4Cy5

Molecular Weight

2616.44 Da

Formulation

Supplied in 20 mM Tris, pH 8.0

Stability & Storage

Store at < -20 °C. Good for 6 months from date of receipt.

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 < -20 °C. Good for 6 months from date of receipt.

For Details:

Related Reagents

Click Chemistry

Enzymes and detection reagents

 

Data Examples

Recombinant Sp Endo-S2 assay using Cy5-G2
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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 37oC. 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.

Specifications

Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Product Datasheets

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Scientific Data

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.

Background: Glycan G2f

Long Name
Glycan G2f
Alternate Names
Glycan G2f

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

  1. Dilute rSp. Endo-S2 to 10 ng/µL in Assay Buffer.
  2. Dilute Cy5-G2f to 0.02 µM in Assay Buffer.
  3. Mix 10 µL of the diluted rSp.Endo-S2 and 10 µL of the Cy5-G2f in a centrifuge tube.
  4. Prepare a negative control by mixing 10 µL of the Cy5-G2f with 10 µL of Assay Buffer.
  5. Incubate the reaction and control at 37o C for 60 minutes.
  6. Stop the reactions and controls by adding 5 uL of 6X gel loading dye to each tube.
  7. 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.
  8. Image the gel with a fluorescent imager for 10 seconds.
  9. 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

FAQs

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