Wheat Germ Agglutinin, AF488 Conjugate

Cat.No.: IC-6201

Product Overview

Wheat Germ Agglutinin (WGA) is one of the most extensively used and studied lectins, specifically binding to N-acetyl-D-glucosamine (GlcNAc) and sialic acid residues. These glycosylated residues are abundantly found on glycoproteins and glycolipids on the cell surface.

This product is a covalent conjugate of WGA and the AF488 fluorescent dye. AF488 is functionally equivalent to Alexa Fluor® 488 in chemical structure, offering similar spectral properties and excellent photostability, resulting in bright, stable green fluorescence. This conjugate is a powerful tool for labeling plasma membranes, neuronal tracing, and localizing glycoconjugates in tissue sections.

Key Features

-Fluorophore: AF488

-Excitation/Emission Maxima: ~499 nm / ~520 nm

-Applications: Fluorescence microscopy, flow cytometry, histochemistry.

-Specificity: Binds to N-acetyl-D-glucosamine and sialic acid residues.

Product Details

Storage Conditions£ºStore frozen at < -15¡æ. Minimize light exposure.

Expiration Date £º12 months from the date of receipt.

Reconstitution Volume £ºRecommended: 0.5 mL of ultrapure water (ddH₂O).

Reconstitution & Storage

1.  Reconstitution:

-Briefly centrifuge the vial before opening to bring the powder to the bottom.

-Add 0.5 mL of ultrapure water and gently vortex until completely dissolved. Avoid excessive foaming.

-This yields a stock solution of approximately 2 mg/mL.

2.  Storage:

-Stock Solution: It is recommended to aliquot the reconstituted stock solution (e.g., 10-20 µL per vial) and store frozen at ¡Ü -15¡æ, protected from light. Avoid repeated freeze-thaw cycles.

-Working Solution: Prepared working solutions should be used immediately and are not recommended for long-term storage.

Spectral Properties

Excitation Wavelength (¦Ëex): 499 nm

Emission Wavelength (¦Ëem): 520 nm

Recommended Filter Set: FITC/GFP filter sets.

Binding Specificity

-WGA primarily recognizes and binds to the following residues:N-acetyl-D-glucosamine (GlcNAc): The primary binding target, with high affinity for chitodextrins (sequences of ¦Â-1,4-linked GlcNAc residues).

-Sialic Acid: An important carbohydrate component on cell surfaces.

-Weak/No Binding: Binds weakly to N-acetylgalactosamine (GalNAc) and does not bind to N-acetylmannosamine (ManNAc).Each WGA monomer contains two identical, non-interacting binding sites.

Application Guide

This product is widely used for labeling glycoconjugates in plasma membranes and the extracellular matrix.

1. Cell Surface Labeling (for Microscopy or Flow Cytometry):Recommended Working Concentration: For most cell types, a starting concentration of 5 - 20 µg/mL is recommended for optimization.

Staining Protocol:

A.Dilute the reconstituted stock solution in an appropriate buffer (e.g., PBS, HBSS) to the working concentration.

B.Incubate cells with the staining solution for 10 - 30 minutes at room temperature (or 4¡ãC), protected from light.

C.Wash cells 2-3 times with buffer to remove unbound conjugate.

  D.Proceed with fluorescence imaging or analysis.

2.  Tissue Staining:

For paraffin-embedded, frozen sections, or fixed tissues, a concentration gradient test is recommended for optimization. Typical incubation times range from 30 minutes to 1 hour, followed by thorough washing.

Note: The optimal working concentration and incubation time may vary depending on the cell type, tissue fixation method, and specific experimental system. A concentration gradient test is recommended for initial use.

Precautions

-Light-Sensitive: Protect from light during reconstitution, dilution, and staining to prevent photobleaching.

-Aliquot Storage: To prevent loss of activity due to repeated freeze-thaw cycles, aliquoting the stock solution is strongly recommended.

-Optimization: The provided protocol is a recommended starting point. Conditions should be optimized for your specific experimental setup (e.g., cell density, instrument parameters).

-Compatibility: Compatible with many fixatives (e.g., paraformaldehyde); however, fixation may alter cell surface glycosylation. Staining after fixation is generally recommended.