L6397

Concanavalin A from Canavalia ensiformis (Jack bean)

peroxidase conjugate, lyophilized powder

• Sinonimo/i: ConA
• Codice UNSPSC: 12352202
• NACRES: NA.32

Proprietà

• Origine biologica: Canavalia ensiformis (jack bean)
• Livello qualitativo: 200
• Coniugato: peroxidase conjugate
• Stato: lyophilized powder
• tecniche: blood typing: suitable; fractionation: suitable
• Impurezze: carbohydrate, essentially free
• Temperatura di conservazione: −20°C

Descrizione

Applicazioni

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

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)

Azioni biochim/fisiol

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.

Confezionamento

Package size based on protein content

Definizione di unità

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

Stato fisico

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

Nota sulla preparazione

Prepared from peroxidase type VI (P8375), using the method of Wilson and Nakane,[1] which promotes conjugation but prevents the interaction between Con A and peroxidase sugar residues.

Risultati analitici

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.

Informazioni sulla sicurezza

• Pittogrammi: GHS08
• Avvertenze: Danger
• Indicazioni di pericolo: H317,H334,H361
• Consigli di prudenza: P202 - P261 - P272 - P280 - P302 + P352 - P308 + P313
• Classi di pericolo: Repr. 2 - Resp. Sens. 1 - Skin Sens. 1
• Codice della classe di stoccaggio: 11 - Combustible Solids
• Classe di pericolosità dell'acqua (WGK): WGK 3
• Punto d’infiammabilità (°F): Not applicable
• Punto d’infiammabilità (°C): Not applicable
• Dispositivi di protezione individuale: Eyeshields, Faceshields, Gloves, type P3 (EN 143) respirator cartridges