iFluor® 700-Wheat Germ Agglutinin (WGA) Conjugate

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Solvent

Water

Spectral properties

Correction Factor (260 nm)	0.09
Correction Factor (280 nm)	0.04
Correction Factor (565 nm)	0.0766
Correction Factor (650 nm)	0.4004
Extinction coefficient (cm ^-1 M ^-1)	220000¹
Excitation (nm)	690
Emission (nm)	713
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)

0.09

Correction Factor (280 nm)

0.04

Correction Factor (565 nm)

0.0766

Correction Factor (650 nm)

0.4004

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

220000¹

Excitation (nm)

690

Emission (nm)

713

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® 700 conjugate of WGA possesses a bright, far-red fluorescence characteristic of the iFluor® 700 dye (Ex/Em = 690/713 nm). Notably, the iFluor® 700 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® 700-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® 700-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® 700-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® 700-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)	0.09
Correction Factor (280 nm)	0.04
Correction Factor (565 nm)	0.0766
Correction Factor (650 nm)	0.4004
Extinction coefficient (cm ^-1 M ^-1)	220000¹
Excitation (nm)	690
Emission (nm)	713
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® 680-Wheat Germ Agglutinin (WGA) Conjugate	684	701	220000¹	0.23¹	0.097	0.094
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® 700-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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Resolving arteriolar wall structures in mouse brain in vivo with three-photon microscopy.
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Journal: Journal of biophotonics (2023): e202200365

A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and Structures, Part III: Reagents for Actin, Tubulin, Cellular Membranes, and Whole Cell and Cytoplasm.
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Evaluation of human platelet granules by structured illumination laser fluorescence microscopy.
Authors: Pluthero, Fred G and Kahr, Walter H A
Journal: Platelets (2023): 2157808

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Authors: Hoffmann, Emily and Gerwing, Mirjam and Niland, Stephan and Niehoff, Rolf and Masthoff, Max and Geyer, Christiane and Wachsmuth, Lydia and Wilken, Enrica and Höltke, Carsten and Heindel, Walter L and Hoerr, Verena and Schinner, Regina and Berger, Philipp and Vogl, Thomas and Eble, Johannes A and Maus, Bastian and Helfen, Anne and Wildgruber, Moritz and Faber, Cornelius
Journal: Journal for immunotherapy of cancer (2023)

Intercomparison of Two Fluorescent Dyes to Visualize Parasitic Fungi (Chytridiomycota) on Phytoplankton.
Authors: Klawonn, Isabell and Dunker, Susanne and Kagami, Maiko and Grossart, Hans-Peter and Van den Wyngaert, Silke
Journal: Microbial ecology (2023): 9-23

LogP of N-acyl-gemcitabine and lectin-corona emerge as key parameters in nanoparticulate intravesical cancer therapy.
Authors: Anzengruber, Maria and Wimmer, Lukas and Szuchar, Raffaela and Skoll, Katharina and Wirth, Michael and Gabor, Franz
Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences (2023): 106330

Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds.
Authors: Pena-Passos, Matheus and Sisti, Laís Soêmis and Mayer, Juliana Lischka Sampaio
Journal: Journal of visualized experiments : JoVE (2022)

A simple and versatile fluorochrome-based procedure for imaging of lipids in arbuscule-containing cells.
Authors: Montero, Héctor and Paszkowski, Uta
Journal: The Plant journal : for cell and molecular biology (2022): 294-301