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L5391

Sigma-Aldrich

Lectin from Bandeiraea simplicifolia (Griffonia simplicifolia)

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

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

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

  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.

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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