Lectin from Bandeiraea simplicifolia (Griffonia simplicifolia)

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: BS-I has a major affinity for terminal α-D-galactosyl residues with a secondary affinity for terminal N-acetyl-α-D-galactosaminyl residues.
• Other Notes: BS-I is a tetrameric lectin consisting of two types of subunits designated A and B. There are five BS-I isolectins with different subunit composition: BSI-B4, BSI-AB3, BSI-A2B2, BSI-A3B and BSI-A4. BSI-B4 is blood group B specific and has an exclusive affinity for terminal α-D-galactosyl residues, whereas BSI-A4 has blood group A specificity and has a major affinity for terminal N-acetyl-α-D-galactosaminyl residues.
• Unit Definition: One unit will form 1 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.
• Physical form: Contains sodium citrate buffer salts and calcium chloride
• Preparation Note: Prepared from peroxidase type VI using a modification of the method of O′Sullivan, et al. Repurified by affinity chromatography after conjugation.
• Analysis Note: Agglutination activity is expressed in μg/mL and is determined from serial dilutions of a 1 mg/mL solution using phosphate buffered saline, pH 6.8, containing, for each lectin, calcium, magnesium, and manganese at different concentrations. This activity is the lowest concentration to agglutinate a 2% suspension of appropriate erythrocytes after 1 hr incubation at 25 °C.