iFluor® 594-Wheat Germ Agglutinin (WGA) Conjugate

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Physical properties

Solvent

Water

Spectral properties

Absorbance (nm)	587
Correction Factor (260 nm)	0.05
Correction Factor (280 nm)	0.04
Extinction coefficient (cm ^-1 M ^-1)	180000¹
Excitation (nm)	588
Emission (nm)	604
Quantum yield	0.53¹

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)

587

Correction Factor (260 nm)

0.05

Correction Factor (280 nm)

0.04

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

180000¹

Excitation (nm)

588

Emission (nm)

604

Quantum yield

0.53¹

Wheat germ agglutinin (WGA) is a lectin that binds to N-acetyl-D-glucosamine and sialic acid. It is of the most studied and useful lectins for it biological applications. Since WGA binds to glycoconjugates its derivatives and conjugates are widely used to label cell membranes and fibrotic scar tissue for fluorescence imaging and analysis. The carbohydrate-binding specificity of WGA is directed against sequences of β-1,4-GlcNAc-linked residues, the chitodextrins. Each monomer contains two identical, non-interacting binding sites which are complementary to 3 or 4 β-1,4-GlcNAc units. Of the monosaccharides examined, only GlcNAc binds to WGA. ManNAc does not bind and GalNAc binds only weakly. iFluor®594 conjugate of WGA might be the brightest red fluorescent WGA conjugate. It exhibits the bright, red fluorescence of the iFluor® 594 dye. iFluor®594 WGA conjugate binds to sialic acid and N-acetylglucosaminyl residues as AF594 WGA conjugate does.

Platform

Fluorescence microscope

Excitation	Cy3/TRITC filter set
Emission	Cy3/TRITC 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® 594-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® 594-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® 594-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 Cy3/TRITC 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® 594-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 Cy3/TRITC filter set.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Absorbance (nm)	587
Correction Factor (260 nm)	0.05
Correction Factor (280 nm)	0.04
Extinction coefficient (cm ^-1 M ^-1)	180000¹
Excitation (nm)	588
Emission (nm)	604
Quantum yield	0.53¹

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® 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® 680-Wheat Germ Agglutinin (WGA) Conjugate	684	701	220000¹	0.23¹	0.097	0.094
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 HeLa cells were stained with iFluor® 594-Wheat Germ Agglutinin (WGA) Conjugate at 5 µg/mL for 30 minutes followed by Hoechst 33342 (AAT Cat# 17535). Image was acquired using fluorescence microscopy using Cy3/TRITC and DAPI filter set.

Citations

View all 1 citations: Citation Explorer

Dihydrolycorine Attenuates Cardiac Fibrosis and Dysfunction by Downregulating Runx1 following Myocardial Infarction
Authors: Ni, Tingjuan and Huang, Xingxiao and Pan, Sunlei and Lu, Zhongqiu
Journal: Oxidative Medicine and Cellular Longevity (2021)

References

View all 50 references: Citation Explorer

Wheat germ agglutinin is involved in the protective action of 24-epibrassinolide on the roots of wheat seedlings under drought conditions.
Authors: Avalbaev, Azamat and Bezrukova, Marina and Allagulova, Chulpan and Lubyanova, Alsu and Kudoyarova, Guzel and Fedorova, Kristina and Maslennikova, Dilara and Yuldashev, Ruslan and Shakirova, Farida
Journal: Plant physiology and biochemistry : PPB (2020): 420-427

Membrane-associated gamma-glutamyl transferase and alkaline phosphatase in the context of concanavalin A- and wheat germ agglutinin-reactive glycans mark seminal prostasome populations from normozoospermic and oligozoospermic men.
Authors: Janković, Tamara and Goč, Sanja and Mitić, Ninoslav and Danilović Luković, Jelena and Janković, Miroslava
Journal: Upsala journal of medical sciences (2020): 10-18

Succinylated Wheat Germ Agglutinin Colocalizes with the Toxoplasma gondii Cyst Wall Glycoprotein CST1.
Authors: Guevara, Rebekah B and Fox, Barbara A and Bzik, David J
Journal: mSphere (2020)

Wheat germ agglutinin is a biomarker of whole grain content in wheat flour and pasta.
Authors: Killilea, David W and McQueen, Rebecca and Abegania, Judi R
Journal: Journal of food science (2020): 808-815

Wheat germ agglutinin liposomes with surface grafted cyclodextrins as bioadhesive dual-drug delivery nanocarriers to treat oral cells.
Authors: Wijetunge, Sashini S and Wen, Jianchuan and Yeh, Chih-Ko and Sun, Yuyu
Journal: Colloids and surfaces. B, Biointerfaces (2020): 110572

Wheat germ agglutinin-conjugated fluorescent pH sensors for visualizing proton fluxes.
Authors: Zhang, Lejie and Zhang, Mei and Bellve, Karl and Fogarty, Kevin E and Castro, Maite A and Brauchi, Sebastian and Kobertz, William R
Journal: The Journal of general physiology (2020)

Bacterial expression, purification and biophysical characterization of wheat germ agglutinin and its four hevein-like domains.
Authors: Leyva, Eduardo and Medrano-Cerano, Jorge L and Cano-Sánchez, Patricia and López-González, Itzel and Gómez-Velasco, Homero and Del Río-Portilla, Federico and García-Hernández, Enrique
Journal: Biopolymers (2019): e23242

Targeted delivery of etoposide, carmustine and doxorubicin to human glioblastoma cells using methoxy poly(ethylene glycol)‑poly(ε‑caprolactone) nanoparticles conjugated with wheat germ agglutinin and folic acid.
Authors: Kuo, Yung-Chih and Chang, Yu-Hsuan and Rajesh, Rajendiran
Journal: Materials science & engineering. C, Materials for biological applications (2019): 114-128

Wheat Germ Agglutinin as a Potential Therapeutic Agent for Leukemia.
Authors: Ryva, Bradley and Zhang, Keman and Asthana, Abhishek and Wong, Derek and Vicioso, Yorleny and Parameswaran, Reshmi
Journal: Frontiers in oncology (2019): 100

Enhanced anti-colon cancer efficacy of 5-fluorouracil by epigallocatechin-3- gallate co-loaded in wheat germ agglutinin-conjugated nanoparticles.
Authors: Wang, Ruoning and Huang, Jinyu and Chen, Jian and Yang, Mengmeng and Wang, Honglan and Qiao, Hongzhi and Chen, Zhipeng and Hu, Lihong and Di, Liuqing and Li, Junsong
Journal: Nanomedicine : nanotechnology, biology, and medicine (2019): 102068