Wheat Germ Agglutinin (WGA)

Wheat germ agglutinin (WGA) is a well-studied lectin known for its binding affinity to N-acetyl-D-glucosamine and sialic acid, making it a valuable tool in various biological applications. Its interaction with glycoconjugates enables widespread use of WGA derivatives and conjugates for fluorescence imaging and analysis, facilitating the labeling of yeast bud scars, fibrotic scar tissue, and the cell membranes of gram bacteria and mammalian cells. WGA specifically targets sequences of β-1,4-GlcNAc-linked residues known as chitodextrins. Each monomer contains two identical, non-interacting binding sites complementary to 3 or 4 β-1,4-GlcNAc units. Among the monosaccharides tested, only GlcNAc shows strong binding to WGA, while ManNAc demonstrates no binding, and GalNAc exhibits weak binding. The mFluor™ Violet 450 labeled WGA is well-excited by the violet laser, emitting a bright blue fluorescence at 445 nm. Notably, the mFluor™ Violet 450 WGA conjugate retains its ability to bind to sialic acid and N-acetylglucosaminyl residues, enhancing its utility in fluorescence imaging and analysis of various scientific investigations.

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

mFluor™ Violet 450-Wheat Germ Agglutinin (WGA) Conjugate stock solution (200X)

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

mFluor™ Violet 450-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

Wash cells twice with a HHBS buffer.
Add 100 µL mFluor™ Violet 450-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 = 406/445 nm.

Fixed Cells Stain

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 450-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 = 406/445 nm.

Spectrum

Open in Advanced Spectrum Viewer

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 500-Wheat Germ Agglutinin (WGA) Conjugate	410	501	25000¹	0.81¹	0.769	0.365
mFluor™ Violet 540-Wheat Germ Agglutinin (WGA) Conjugate	402	535	18000¹	0.21¹	1.326	0.543

References

View all 50 references: Citation Explorer

Diverse Enterococcus faecalis strains show heterogeneity in biofilm properties.
Authors: Schaffer, Scott D and Hutchison, Carissa A and Rouchon, Candace N and Mdluli, Nontokozo V and Weinstein, Arielle J and McDaniel, Dennis and Frank, Kristi L
Journal: Research in microbiology (2023): 103986

Expansion Microscopy of Bacillus subtilis.
Authors: Middelhauve, Viola and Siebrasse, Jan Peter and Kubitscheck, Ulrich
Journal: Methods in molecular biology (Clifton, N.J.) (2023): 191-202

Human serum albumin nanoparticles as a versatile vehicle for targeted delivery of antibiotics to combat bacterial infections.
Authors: Skoll, Katharina and Palmetzhofer, Julia and Lummerstorfer, Maria and Anzengruber, Maria and Gabor, Franz and Wirth, Michael
Journal: Nanomedicine : nanotechnology, biology, and medicine (2023): 102685

Microencapsulation of Probiotics with Soy Protein Isolate and Alginate for the Poultry Industry.
Authors: Babot, Jaime D and Argañaraz-Martínez, Eloy and Apella, María C and Perez Chaia, Adriana
Journal: Food and bioprocess technology (2023): 1478-1487

Ultrasensitive Fluorescence Lateral Flow Assay for Simultaneous Detection of Pseudomonas aeruginosa and Salmonella typhimurium via Wheat Germ Agglutinin-Functionalized Magnetic Quantum Dot Nanoprobe.
Authors: Tu, Zhijie and Yang, Xingsheng and Dong, Hao and Yu, Qing and Zheng, Shuai and Cheng, Xiaodan and Wang, Chongwen and Rong, Zhen and Wang, Shengqi
Journal: Biosensors (2022)

Page updated on October 4, 2024

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Physical properties

Solvent

Water

Spectral properties

Absorbance (nm)	406
Correction Factor (260 nm)	0.338
Correction Factor (280 nm)	0.078
Extinction coefficient (cm ^-1 M ^-1)	35000¹
Excitation (nm)	406
Emission (nm)	445
Quantum yield	0.81¹

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501