Lectin from Ulex europaeus (gorse, furze), peroxidase conjugate, lyophilized powder

Stock Code: 3587102
Manufacturer Part No: L8146-.5MG

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Descriptions

Analysis Note

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.

Application

General Western Blot Protocol: Glycoprotein sample size: 500ng Lectin Concentration: 0.1ug/mlLoad 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

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.

Linkage

Conjugates are prepared from affinity purified lectin.

Packaging

0.5, 1 mg in serum bottle

Package size based on protein content.

Physical form

Contains sodium citrate buffer salts

Preparation Note

Prepared from peroxidase, Type VI (P8375), using the two-step glutaraldehyde method of Avrameas and Ternyck.Repurified by affinity chromatography after conjugation.

Unit Definition

One unit will form 1.0?mg purpurogallin in 20?sec from pyrogallol at pH?6.0 at 20?°C.