LECTIN FROM TRITICUM VULGARIS PEROXIDASE LABELED (C005B-466058)

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.

Unit Definition

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

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.