Not available outside of the UK & Ireland.

Analysis Note

Where reported, agglutination activity is expressed in µg/ml and is determined from serial dilutions in phosphate buffered saline, pH 6.8, containing Ca²⁺ and Mn²⁺ of a 1 mg per mL solution. This activity is the lowest concentration to agglutinate a 2% suspension of human erythrocytes after 1 hr 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)

Detects glycoproteins containing α-D-mannose, α-D-glucose when used with appropriate peroxidase substrate

Biochem/physiol Actions

Con A is not blood group specific but has an affinity for terminal α-D-mannosyl and α-D-glucosyl residues. Ca²⁺ and Mn²⁺ ions are required for activity. Con A dissociates into dimers at pH 5.6 or below. Between pH 5.8 and pH 7.0, Con A exists as a tetramer; above pH 7.0 higher aggregates are formed. Con A exhibits mitogenic activity which is dependent on its degree of aggregation. Succinylation results in an active dimeric form which remains a dimer above pH 5.6.

Packaging

Package size based on protein content

Physical form

Lyophilized powder containing tris-citrate buffer salts and trace calcium and manganese

Preparation Note

Prepared from peroxidase type VI (P8375), using the method of Wilson and Nakane, which promotes conjugation but prevents the interaction between Con A and peroxidase sugar residues.

Unit Definition

One unit will form 1 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.