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AAT Bioquest

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OverviewpdfSDSpdfProtocol


Example protocol


PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

AF488-Wheat Germ Agglutinin (WGA) Conjugate stock solution (200X)
Add 500 µL of ddH2O into the powder form to make 2 mg/mL stock solution.
Note     The reconstituted conjugate solution can be stored at 2-8 °C for short-term storage or at -20 °C for long-term storage.

PREPARATION OF WORKING SOLUTION

AF488-Wheat Germ Agglutinin (WGA) Conjugate working solution (1X)
Add 5 µL of 200X WGA conjugate solution to 1 mL HHBS Buffer.
Note     The optimized staining concentration may be different with different cell lines. The recommended starting concentration is 5-10 µg/mL for live cells.

SAMPLE EXPERIMENTAL PROTOCOL

Warm the vial to room temperature centrifuge briefly before opening. Staining protocols vary with applications. Appropriate dilution of conjugates should be determined experimentally.

Live Cells Stain
  1. Wash cells twice with a HHBS buffer.
  2. Add 100 µL AF488-WGA working solution.
  3. Incubate cells with WGA working solution for 10-30 minutes at 37 °C.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using FITC filter set. 

Fixed Cells Stain
WGA conjugates can be also used to stain fixed cells.
  1. Fix cells with 4% Formaldehyde in PBS.
    Note     For fixed cell membrane staining, it is recommended to stain without permeabilization step. Permeabilized step can after fixation will lead to intracellular compartments stain such as Golgi and Endoplasmic Reticulum (ER) structures.
  2. Add 100 µL AF488-WGA working solution.
  3. Incubate cells with WGA working solution for 10-30 minutes at room temperature.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using FITC filter set.