iFluor® 350-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)	0.83
Correction Factor (280 nm)	0.23
Extinction coefficient (cm ^-1 M ^-1)	20000¹
Excitation (nm)	345
Emission (nm)	450
Quantum yield	0.95¹

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)

0.83

Correction Factor (280 nm)

0.23

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

20000¹

Excitation (nm)

345

Emission (nm)

450

Quantum yield

0.95¹

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® 350 conjugate of WGA possesses a bright, blue fluorescence characteristic of the iFluor® 350 dye (Ex/Em = 345/450 nm). Notably, the iFluor® 350 WGA conjugate, like Alexa Fluor® 350 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	DAPI filter set
Emission	DAPI 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® 350-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® 350-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® 350-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 DAPI 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® 350-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 DAPI filter set.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Correction Factor (260 nm)	0.83
Correction Factor (280 nm)	0.23
Extinction coefficient (cm ^-1 M ^-1)	20000¹
Excitation (nm)	345
Emission (nm)	450
Quantum yield	0.95¹

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	587	603	200000¹	0.53¹	0.05	0.04
iFluor® 647-Wheat Germ Agglutinin (WGA) Conjugate	656	670	250000¹	0.25¹	0.03	0.03
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 and fixed HeLa cells were stained with iFluor® 350-Wheat Germ Agglutinin (WGA) Conjugate at 10 µg/mL for 30 minutes. The image was acquired on a fluorescence microscope using a DAPI filter set.

References

View all 10 references: Citation Explorer

Chairside molecular imaging of aberrant glycosylation in subjects with suspicious oral lesions using fluorescently labeled wheat germ agglutinin.
Authors: Baeten, John and Johnson, Alexander and Sunny, Sumsum and Suresh, Amritha and Birur, Praveen and Uma, K and Kademani, Deepak
Journal: Head & neck (2018): 292-301

Chitin distribution in the Oithona digestive and reproductive systems revealed by fluorescence microscopy.
Authors: Sugier, Kevin and Vacherie, Benoit and Cornils, Astrid and Wincker, Patrick and Jamet, Jean-Louis and Madoui, Mohammed-Amin
Journal: PeerJ (2018): e4685

Antifungal curcumin promotes chitin accumulation associated with decreased virulence of Sporothrix schenckii.
Authors: Huang, Lilin and Zhang, Jing and Song, Tianzhang and Yuan, Liyan and Zhou, Junjie and Yin, Hongling and He, Tailong and Gao, Wenchao and Sun, Yao and Hu, Xuchu and Huang, Huaiqiu
Journal: International immunopharmacology (2016): 263-270

Endothelial glycocalyx structure in the intact carotid artery: a two-photon laser scanning microscopy study.
Authors: Reitsma, Sietze and oude Egbrink, Mirjam G A and Vink, Hans and van den Berg, Bernard M and Passos, Valéria Lima and Engels, Wim and Slaaf, Dick W and van Zandvoort, Marc A M J
Journal: Journal of vascular research (2011): 297-306

Isolation and characterization of Saccharomyces cerevisiae mutants resistant to Calcofluor white.
Authors: Roncero, C and Valdivieso, M H and Ribas, J C and Durán, A
Journal: Journal of bacteriology (1988): 1950-4

The binding of monosaccharides to wheat germ agglutinin: fluorescence and NMR investigations.
Authors: Midoux, P and Grivet, J P and Delmotte, F and Monsigny, M
Journal: Biochemical and biophysical research communications (1984): 603-11

Fluorescence quenching of tryptophan by trifluoroacetamide.
Authors: Midoux, P and Wahl, P and Auchet, J C and Monsigny, M
Journal: Biochimica et biophysica acta (1984): 16-25

Isolation of murine pluripotent hemopoietic stem cells.
Authors: Visser, J W and Bauman, J G and Mulder, A H and Eliason, J F and de Leeuw, A M
Journal: The Journal of experimental medicine (1984): 1576-90

Localization of chitin in the cell wall of Candida albicans by means of wheat germ agglutinin. Fluorescence and ultrastructural studies.
Authors: Tronchin, G and Poulain, D and Herbaut, J and Biguet, J
Journal: European journal of cell biology (1981): 121-8

Nanosecond-pulse fluorimetry of wheat-germ agglutinin (lectin).
Authors: Privat, J P and Wahl, P and Monsigny, M and Auchet, J C
Journal: European journal of biochemistry (1976): 573-80