Wheat Germ Agglutinin (WGA)
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
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.
PREPARATION OF WORKING SOLUTION
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.
- Wash cells twice with a HHBS buffer.
Add 100 µL mFluor™ Violet 500-WGA working solution.
- Incubate cells with WGA working solution for 10-30 minutes at 37 °C.
- Wash cells twice with HHBS buffer.
Image cells on a fluorescence microscope using Ex/Em = 410/501 nm.
WGA conjugates can be also used to stain fixed cells.
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.Add 100 µL mFluor™ Violet 500-WGA working solution.
- Incubate cells with WGA working solution for 10-30 minutes at room temperature.
- Wash cells twice with HHBS buffer.
Image cells on a fluorescence microscope using Ex/Em = 410/501 nm.
Spectrum
Product family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
mFluor™ Violet 450-Wheat Germ Agglutinin (WGA) Conjugate | 406 | 445 | 350001 | 0.811 | 0.338 | 0.078 |
mFluor™ Violet 540-Wheat Germ Agglutinin (WGA) Conjugate | 402 | 535 | 180001 | 0.211 | 1.326 | 0.543 |
References
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