L5391
Lectin from Bandeiraea simplicifolia (Griffonia simplicifolia)
Isolectin B4 (BSI-B4), peroxidase conjugate, lyophilized powder
Synonym(s):
Bandeirea simplicifolia agglutinin, BS-I
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About This Item
UNSPSC Code:
12352202
NACRES:
NA.32
Recommended Products
biological source
Bandieraea simplicifolia
Quality Level
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
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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
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.
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
Certificates of Analysis (COA)
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