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Cy5-Fuc Labeled Glycan G2f

Catalog # GL302 | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Key Benefits:

Learn more about Fluorescent Glycan Labeling and Detection

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Setting the standard in quality research reagents

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

Product Documents for Cy5-Fuc Labeled Glycan G2f

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.