L3892

Lectin from Triticum vulgaris (wheat)

peroxidase conjugate, lyophilized powder

• Synonym(s): WGA, Wheat germ agglutinin
• NACRES: NA.32
• UNSPSC Code: 12352202
• Form: lyophilized powder
• Assay: ≥85% protein basis (Warburg-Christian)
• Biological source: Triticum vulgaris

Properties

• biological source: Triticum vulgaris
• Quality Level: 200
• conjugate: peroxidase conjugate
• assay: ≥85% protein basis (Warburg-Christian)
• form: lyophilized powder
• potency: <40 μg/mL agglutination activity
• peroxidase activity: 50-200 units/mg protein
• composition: Protein, ~90% modified Warburg-Christian
• storage temp.: −20°C

Description

Application

Detects glycoproteins containing β(1→4)-N-acetyl-D-glucosamine when used with appropriate peroxidase substrate

General Western Blot Protocol:

• Glycoprotein sample size: 500ng
• Lectin Concentration: 0.1ug/ml

1. Load samples at 500 ng of glycoprotein per lane
2. Run 4-20% Bis-Tris SDS page gel
3. Transfer gel to a PVDF membrane
4. Block membrane for 1 hr at RT with RIPA buffer (R0278 Sigma)
5. Incubate HRP lectin at 0.1ug/ml with RIPA buffer for 2 hours at RT
6. Wash membrane 5 x 5 minutes with 25ml RIPA buffer
7. Detect using chemiluminescent substrate (CPS1-120)

Biochem/physiol Actions

WGA is not blood group specific but has an affinity for N-acetyl-β-D-glucosaminyl residues and N-acetyl-β-D-glucosamine oligomers. WGA contains no protein-bound carbohydrate.

Packaging

Package size based on protein content.

Physical form

Contains citrate buffer

Preparation Note

Conjugates are prepared from affinity purified lectin.

Prepared from peroxidase type VI (P8375) using a modification of the method of O′Sullivan,[1] which favors low molecular weight conjugates. Repurified after conjugation by affinity chromatography.

Other Notes

One unit will form 1.0 mg of purpurogallin from pyrogallol in 20 sec at pH 6.0 at 20 °C.

Safety Information

• Storage Class: 11 - Combustible Solids
• wgk: WGK 3
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: Eyeshields, Gloves, type N95 (US)