Wheat Germ Agglutinin (WGA)

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

1. Add 500 µL of ddH₂O into the powder form to make a 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.

1. 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.

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

1. Wash cells twice with a HHBS buffer.

2. Add 100 µL iFluor® 532-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 Cy3/TRITC filter set.

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 the permeabilization step. A permeabilization step after fixation can facilitate staining intracellular compartments such as Golgi and Endoplasmic Reticulum (ER) structures.

2. Add 100 µL iFluor® 532-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 Cy3/TRITC filter set.