L6397
Concanavalin A from Canavalia ensiformis (Jack bean)
peroxidase conjugate, lyophilized powder
Synonym(s):
ConA
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About This Item
UNSPSC Code:
12352202
NACRES:
NA.32
Recommended Products
biological source
Canavalia ensiformis (jack bean)
Quality Level
conjugate
peroxidase conjugate
form
lyophilized powder
technique(s)
blood typing: suitable
fractionation: suitable
impurities
carbohydrate, essentially free
storage temp.
−20°C
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Application
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
- 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
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
Unit Definition
One unit will form 1 mg purpurogallin in 20 sec from pyrogallol at pH 6.0 at 20 °C.
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.
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.
signalword
Danger
hcodes
Hazard Classifications
Repr. 2 - Resp. Sens. 1 - Skin Sens. 1
Storage Class
11 - Combustible Solids
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Faceshields, Gloves, type P3 (EN 143) respirator cartridges
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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