mFluor™ Violet 450-Wheat Germ Agglutinin (WGA) Conjugate

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
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Shipping	Standard overnight for United States, inquire for international

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

Overview SDS Protocol

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

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.

Platform

Fluorescence microscope

Excitation	406 nm
Emission	445 nm
Recommended plate	Black wall/clear bottom

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

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

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

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Human serum albumin nanoparticles as a versatile vehicle for targeted delivery of antibiotics to combat bacterial infections.
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Journal: Methods in molecular biology (Clifton, N.J.) (2023): 191-202

Ultrasensitive Fluorescence Lateral Flow Assay for Simultaneous Detection of Pseudomonas aeruginosa and Salmonella typhimurium via Wheat Germ Agglutinin-Functionalized Magnetic Quantum Dot Nanoprobe.
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Journal: Biosensors (2022)

Universal and ultrasensitive detection of foodborne bacteria on a lateral flow assay strip by using wheat germ agglutinin-modified magnetic SERS nanotags.
Authors: Tu, Zhijie and Cheng, Siyun and Dong, Hao and Wang, Wenqi and Yang, Xingsheng and Gu, Bing and Wang, Shengqi and Wang, Chongwen
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Assay for Assessing Mucin Binding to Bacteria and Bacterial Proteins.
Authors: Grigoryeva, Lubov S and Rehman, Saima and White, Richard C and Garnett, James A and Cianciotto, Nicholas P
Journal: Bio-protocol (2021): e3933

Degradation of Wheat Germ Agglutinin during Sourdough Fermentation.
Authors: Tovar, Luis E Rojas and Gänzle, Michael G
Journal: Foods (Basel, Switzerland) (2021)

An efficient SERS platform for the ultrasensitive detection of Staphylococcus aureus and Listeria monocytogenes via wheat germ agglutinin-modified magnetic SERS substrate and streptavidin/aptamer co-functionalized SERS tags.
Authors: Cheng, Siyun and Tu, Zhijie and Zheng, Shuai and Cheng, Xiaodan and Han, Han and Wang, Chongwen and Xiao, Rui and Gu, Bing
Journal: Analytica chimica acta (2021): 339155

Protection of the intestinal epithelium of poultry against deleterious effects of dietary lectins by a multi-strain bacterial supplement.
Authors: Babot, Jaime Daniel and Argañaraz-Martínez, Eloy and Quiroga, María and Grande, Sonia María and Apella, María Cristina and Perez Chaia, Adriana
Journal: Research in veterinary science (2021): 27-35