Merck, L3892-2MG, Lectin from Triticum vulgaris (wheat)

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

Load 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
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, 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.