iFluor® 680-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

Correction Factor (260 nm)	0.097
Correction Factor (280 nm)	0.094
Extinction coefficient (cm ^-1 M ^-1)	220000¹
Excitation (nm)	684
Emission (nm)	701
Quantum yield	0.23¹

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

Correction Factor (260 nm)

0.097

Correction Factor (280 nm)

0.094

Extinction coefficient (cm ^-1 M ^-1)

220000¹

Excitation (nm)

684

Emission (nm)

701

Quantum yield

0.23¹

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 iFluor® 680 conjugate of WGA possesses a bright, far-red fluorescence characteristic of the iFluor® 680 dye (Ex/Em = 684/701 nm). Notably, the iFluor® 680 WGA conjugate, like Alexa Fluor® 680 WGA conjugate, retains its ability to bind to sialic acid and N-acetylglucosaminyl residues, enhancing its utility in fluorescence imaging and analysis for various scientific investigations.

Platform

Fluorescence microscope

Excitation	Cy5 filter set
Emission	Cy5 filter set
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

iFluor® 680-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

iFluor® 680-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 iFluor® 680-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 Cy5 filter set.

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 iFluor® 680-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 Cy5 filter set.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Correction Factor (260 nm)	0.097
Correction Factor (280 nm)	0.094
Extinction coefficient (cm ^-1 M ^-1)	220000¹
Excitation (nm)	684
Emission (nm)	701
Quantum yield	0.23¹

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (260 nm)	Correction Factor (280 nm)
iFluor® 488-Wheat Germ Agglutinin (WGA) Conjugate	491	516	75000¹	0.9¹	0.21	0.11
iFluor® 555-Wheat Germ Agglutinin (WGA) Conjugate	557	570	100000¹	0.64¹	0.23	0.14
iFluor® 594-Wheat Germ Agglutinin (WGA) Conjugate	588	604	180000¹	0.53¹	0.05	0.04
iFluor® 647-Wheat Germ Agglutinin (WGA) Conjugate	656	670	250000¹	0.25¹	0.03	0.03
iFluor® 350-Wheat Germ Agglutinin (WGA) Conjugate	345	450	20000¹	0.95¹	0.83	0.23
iFluor® 532-Wheat Germ Agglutinin (WGA) Conjugate	537	560	90000¹	0.68¹	0.26	0.16
iFluor® 700-Wheat Germ Agglutinin (WGA) Conjugate	690	713	220000¹	0.23¹	0.09	0.04
iFluor® 750-Wheat Germ Agglutinin (WGA) Conjugate	757	779	275000¹	0.12¹	0.044	0.039
iFluor® 790-Wheat Germ Agglutinin (WGA) Conjugate	787	812	250000¹	0.13¹	0.1	0.09

Images

Figure 1. Live and fixed HeLa cells were stained with iFluor® 680-Wheat Germ Agglutinin (WGA) Conjugate at 10 µg/mL for 30 minutes. The image was acquired on a fluorescence microscope using a Cy5 filter set.

References

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Metabolic reprogramming and flux to cell envelope precursors in a pentose phosphate pathway mutant increases MRSA resistance to β-lactam antibiotics.
Authors: Zeden, Merve S and Gallagher, Laura A and Bueno, Emilio and Nolan, Aaron C and Ahn, Jongsam and Shinde, Dhananjay and Razvi, Fareha and Sladek, Margaret and Burke, Órla and O'Neill, Eoghan and Fey, Paul D and Cava, Felipe and Thomas, Vinai C and O'Gara, James P
Journal: bioRxiv : the preprint server for biology (2023)

Surface-Templated Glycopolymer Nanopatterns Transferred to Hydrogels for Designed Multivalent Carbohydrate-Lectin Interactions across Length Scales.
Authors: Singh, Anamika and Arango, Juan C and Shi, Anni and d'Aliberti, Joseph B and Claridge, Shelley A
Journal: Journal of the American Chemical Society (2023): 1668-1677

Wheat Germ Agglutinin Affinity Chromatography Enrichment and Glyco-Proteomic Characterization of Tetrodotoxin-Binding Proteins from the Plasma of Cultured Tiger Pufferfish (Takifugu rubripes).
Authors: Zhang, Yafei and Minami, Ryoma and Tatsuno, Ryohei and Gao, Wei and Ueno, Mikinori and Yamada, Akinori and Yoshida, Asami and Sedanza, Mary Grace and Arima, Kazunari and Takatani, Tomohiro and Yamaguchi, Kenichi and Oshima, Yuji and Arakawa, Osamu
Journal: Bioscience, biotechnology, and biochemistry (2023)

Dose-dependent Effects of PRC2 and HDAC Inhibitors on Cardiomyocyte Hypertrophy Induced by Phenylephrine.
Authors: Zhao, Zhenyi and Lv, Jian and Guo, Ningning and Guo, Qiuxiao and Zeng, Sai and Fang, Yu and Chen, Weixin and Wang, Zhihua
Journal: Current drug targets (2023): 371-378

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

Synthetic assembly of a series of glycopolymers having sialyl α2-3 lactose moieties connected with longer spacer arms.
Authors: Adachi, Ryota and Matsushita, Takahiko and Koyama, Tetsuo and Hatano, Ken and Matsuoka, Koji
Journal: Bioorganic & medicinal chemistry (2023): 117209

Resolving arteriolar wall structures in mouse brain in vivo with three-photon microscopy.
Authors: Qin, Mengyuan and Huang, Jie and Zhong, Jincheng and Zhang, Yingxian and Tong, Shen and Cheng, Hui and Deng, Xiangquan and Zheng, Lei and Zhang, Wanjian and Qiu, Ping and Wang, Ke
Journal: Journal of biophotonics (2023): e202200365

The counterbalance of endothelial glycocalyx and high wall shear stress to low-density lipoprotein concentration polarization in mouse ear skin arterioles.
Authors: Deng, Zhilan and Sun, Kaixin and Sha, Dongyu and Zhang, Yinuo and Guo, Jiaxin and Yan, Guiqin and Zhang, Weichen and Liu, Ming and Deng, Xiaoyan and Kang, Hongyan and Sun, Anqiang
Journal: Atherosclerosis (2023): 24-33

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

Exosomes and Microvesicles From Adipose-derived Mesenchymal Stem Cells Protects the Endothelial Glycocalyx From LPS Injury.
Authors: Taghavi, Sharven and Abdullah, Sarah and Shaheen, Farhana and Packer, Jacob and Duchesne, Juan and Braun, Stephen E and Steele, Chad and Pociask, Derek and Kolls, Jay and Jackson-Weaver, Olan
Journal: Shock (Augusta, Ga.) (2023)