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

1 mg in glass bottle

Package size based on protein content.

2 mg in serum bottle

Physical form

Contains citrate buffer

Preparation Note

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

Conjugates are prepared from affinity purified lectin.

Unit Definition

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