Description_x000D_ Packaging_x000D_ 0.5, 1 mg in serum bottle_x000D_ Application_x000D_ General Western Blot Protocol:_x000D_ • Glycoprotein sample size: 500ng_x000D_ • Lectin Concentration: 0.1ug/ml_x000D_ Load samples at 500 ng of glycoprotein per lane_x000D_ • Run 4-20% Bis-Tris SDS page gel_x000D_ • Transfer gel to a PVDF membrane_x000D_ • Block membrane for 1 hr at RT with RIPA buffer (R0278 Sigma)_x000D_ • Incubate HRP lectin at 0.1ug/ml with RIPA buffer for 2 hours at RT_x000D_ • Wash membrane 5 x 5 minutes with 25ml RIPA buffer_x000D_ • Detect using chemiluminescent substrate (CPS1-120)_x000D_ Biochem/physiol Actions_x000D_ 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._x000D_ Packaging_x000D_ Package size based on protein content._x000D_ Linkage_x000D_ Conjugates are prepared from affinity purified lectin._x000D_ Unit Definition_x000D_ One unit will form 1.0 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C._x000D_ Physical form_x000D_ Contains sodium citrate buffer salts_x000D_ Preparation Note_x000D_ Prepared from peroxidase, Type VI (P8375), using the two-step glutaraldehyde method of Avrameas and Ternyck._x000D_ Repurified by affinity chromatography after conjugation._x000D_ Analysis Note_x000D_ 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.