L5391

Lectin from Bandeiraea simplicifolia (Griffonia simplicifolia)

Isolectin B₄ (BSI-B₄), peroxidase conjugate, lyophilized powder

• Synonym(s): Bandeirea simplicifolia agglutinin, BS-I
• UNSPSC Code: 12352202
• NACRES: NA.32

Properties

• biological source: Bandieraea simplicifolia
• Quality Level: 200
• conjugate: peroxidase conjugate
• Assay: ≥90% protein basis (Warburg-Christian)
• form: lyophilized powder
• potency: <200 μg per mL agglutination activity (using human blood group B erythrocytes)
• peroxidase activity: 40-160 units/mg protein
• composition: Protein, ~95% modified Warburg-Christian
• technique(s): blood typing: suitable; fractionation: suitable
• storage temp.: −20°C

Description

Application

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

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.

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves