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Wheat Germ Agglutinin (WGA)

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.

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)
  1. 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)
  1. 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
  1. Wash cells twice with a HHBS buffer.
  2. Add 100 µL iFluor® 680-WGA working solution.

  3. Incubate cells with WGA working solution for 10-30 minutes at 37 °C.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using Cy5 filter set.

Fixed Cells Stain

WGA conjugates can be also used to stain fixed cells.

  1. 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.

  2. Add 100 µL iFluor® 680-WGA working solution.

  3. Incubate cells with WGA working solution for 10-30 minutes at room temperature.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using Cy5 filter set.

Spectrum

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
iFluor® 488-Wheat Germ Agglutinin (WGA) Conjugate4915167500010.910.210.11
iFluor® 555-Wheat Germ Agglutinin (WGA) Conjugate55757010000010.6410.230.14
iFluor® 594-Wheat Germ Agglutinin (WGA) Conjugate58760320000010.5310.050.04
iFluor® 647-Wheat Germ Agglutinin (WGA) Conjugate65667025000010.2510.030.03
iFluor® 350-Wheat Germ Agglutinin (WGA) Conjugate3454502000010.9510.830.23
iFluor® 532-Wheat Germ Agglutinin (WGA) Conjugate5375609000010.6810.260.16
iFluor® 700-Wheat Germ Agglutinin (WGA) Conjugate69071322000010.2310.090.04
iFluor® 750-Wheat Germ Agglutinin (WGA) Conjugate75777927500010.1210.0440.039
iFluor® 790-Wheat Germ Agglutinin (WGA) Conjugate78781225000010.1310.10.09

References

View all 50 references: Citation Explorer
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
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
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
Page updated on October 4, 2024

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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-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12171501

Platform

Fluorescence microscope

ExcitationCy5 filter set
EmissionCy5 filter set
Recommended plateBlack wall, clear bottom
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.
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.
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.