mFluor™ Violet 540-Wheat Germ Agglutinin (WGA) Conjugate

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Telephone	1-800-990-8053
Fax	1-800-609-2943
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Physical properties

Solvent

Water

Spectral properties

Absorbance (nm)	401
Correction Factor (260 nm)	1.326
Correction Factor (280 nm)	0.543
Extinction coefficient (cm ^-1 M ^-1)	18000¹
Excitation (nm)	402
Emission (nm)	535
Quantum yield	0.21¹

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)

401

Correction Factor (260 nm)

1.326

Correction Factor (280 nm)

0.543

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

18000¹

Excitation (nm)

402

Emission (nm)

535

Quantum yield

0.21¹

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 540 labeled WGA is well-excited by the violet laser, emitting a bright green fluorescence at 535 nm. Notably, the mFluor™ Violet 540 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	402 nm
Emission	535 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 540-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 540-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 540-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 = 402/535 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 540-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 = 402/535 nm.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Absorbance (nm)	401
Correction Factor (260 nm)	1.326
Correction Factor (280 nm)	0.543
Extinction coefficient (cm ^-1 M ^-1)	18000¹
Excitation (nm)	402
Emission (nm)	535
Quantum yield	0.21¹

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 450-Wheat Germ Agglutinin (WGA) Conjugate	406	445	35000¹	0.81¹	0.338	0.078
mFluor™ Violet 500-Wheat Germ Agglutinin (WGA) Conjugate	410	501	25000¹	0.81¹	0.769	0.365

References

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Application of immuno- and affinity labeling with fluorescent dyes to in-resin CLEM of Epon-embedded cells.
Authors: Tanida, Isei and Yamaguchi, Junji and Suzuki, Chigure and Kakuta, Soichiro and Uchiyama, Yasuo
Journal: Heliyon (2023): e17394

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

Supramolecular Binding with Lectins: A New Route for Non-Covalent Functionalization of Polysaccharide Matrices.
Authors: Montroni, Devis and Di Giosia, Matteo and Calvaresi, Matteo and Falini, Giuseppe
Journal: Molecules (Basel, Switzerland) (2022)

Fluorescence background quenching as a means to increase Signal to Background ratio - a proof of concept during Nerve Imaging.
Authors: Buckle, Tessa and van der Wal, Steffen and van Willigen, Danny M and Aalderink, Germaine and KleinJan, Gijs H and van Leeuwen, Fijs W B
Journal: Theranostics (2020): 9890-9898

Cell Envelope Integrity and Capsule Characterization of Rhodotorula mucilaginosa Strains from Clinical and Environmental Sources.
Authors: Yockey, Johnathan and Andres, Luke and Carson, Moleigh and Ory, Jeramia J and Reese, Amy J
Journal: mSphere (2019)

Detection of white head symptoms of panicle blast caused by Pyricularia oryzae using cut-flower dye.
Authors: Hayashi, Keiko and Yoshida, Tomofumi and Hayano-Saito, Yuriko
Journal: Plant methods (2019): 159

Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis.
Authors: de Wit, Sanne and Zeune, Leonie L and Hiltermann, T Jeroen N and Groen, Harry J M and Dalum, Guus van and Terstappen, Leon W M M
Journal: Cancers (2018)

Poly - (l) - glutamic acid drug delivery system for the intravesical therapy of bladder cancer using WGA as targeting moiety.
Authors: Apfelthaler, C and Anzengruber, M and Gabor, F and Wirth, M
Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenst (2017): 131-139

Pathogenic features and characteristics of food borne pathogens biofilm: Biomass, viability and matrix.
Authors: Lin, Shiqi and Yang, Ling and Chen, Gu and Li, Bing and Chen, Dingqiang and Li, Lin and Xu, Zhenbo
Journal: Microbial pathogenesis (2017): 285-291

Quantification, 2DE analysis and identification of enriched glycosylated proteins from mouse muscles: Difficulties and alternatives.
Authors: de Fátima MenegociEugênio, Patrícia and Assunção, Nilson Antonio and Sciandra, Francesca and Aquino, Adriano and Brancaccio, Andrea and Carrilho, Emanuel
Journal: Electrophoresis (2016): 321-34