Lectin from Ulex europaeus (gorse, furze)

Description

• Application: General Western Blot Protocol:Glycoprotein sample size: 500ngLectin Concentration: 0.1ug/mlLoad samples at 500 ng of glycoprotein per laneRun 4-20% Bis-Tris SDS page gelTransfer gel to a PVDF membraneBlock 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 RTWash membrane 5 x 5 minutes with 25ml RIPA bufferDetect 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.
• Linkage: Conjugates are prepared from affinity purified lectin.
• Unit Definition: One unit will form 1.0 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.
• 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.[1]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.