Merck, L8146-.5MG, Lectin from Ulex europaeus (gorse, furze)

Application
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
Ulex europaeus agglutinin has anti-H blood group specificity. There are two types of lectin: UEA I has an affinity for L-fucose and UEA II has an affinity for N,N′-diacetylchitobiose. The mol. wt. of UEA I was initially found to be 170,000 daltons. Later reports indicate that UEA I may form aggregates at neutral and basic pH and that the correct mol. wt. is 68,000 daltons.
Packaging
Package size based on protein content.
Physical form
Contains sodium citrate buffer salts
Preparation Note
Prepared from peroxidase, Type VI (P8375), using the two-step glutaraldehyde method of Avrameas and Ternyck.
Repurified by affinity chromatography after conjugation.
Analysis Note
Agglutination activity is expressed in μg/ml and is determined from serial dilutions in phosphate buffered saline, pH 6.8, of a 1 mg/ml solution. This activity is the lowest concentration to agglutinate a 2% suspension of human blood group O erythrocytes after 1 hour incubation at 25 °C.
Other Notes
Conjugates are prepared from affinity purified lectin.
One unit will form 1.0 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.