Cy5-Fuc Labeled Glycan G2f

R&D Systems | Catalog # GL302

R&D Systems
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Key Product Details

Key Benefits

Learn more about Fluorescent Glycan Labeling and Detection

Conjugate

Cy5

Product Summary for Cy5-Fuc Labeled Glycan G2f

Cy5 Labeled Antibody Glycan G2

Formula

Fuc1Gal2Man3GlcNAc4Cy5

Molecular Weight

2616.44 Da

Formulation

Supplied in 20 mM Tris, pH 8.0

Stability & Storage

• 6 months from date of receipt, -20 to -70 °C
as supplied.
• 3 months, -20 to -70 °C under sterile
conditions after opening.
• Use a manual defrost freezer and avoid
repeated freeze-thaw cycles.

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
View Larger Image

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.

Scientific Data Images 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.

Formulation, Preparation, and Storage

Formulation

No With Carrier Product Bottling Info

Reconstitution Instructions

Reconstitute at 100 μg/mL in PBS containing at least 0.1% human or bovine serum albumin.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Glycan G2f

Long Name

Glycan G2f

Additional Glycan G2f Products

Product Documents for Cy5-Fuc Labeled Glycan G2f

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Cy5-Fuc Labeled Glycan G2f

For research use only

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Protocols

View specific protocols for Cy5-Fuc Labeled Glycan G2f (GL302):

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