Although many buffers can be employed for reconstituting and diluting this lectin, 10 mM HEPES buffered saline, pH 8.5, 0.1 mM CaCl2 is recommended. For preserving solutions stored at 4 ºC, 0.08% sodium azide can be used.
Con A recognizes α-linked mannose present as part of a core oligosaccharide in many serum and membrane glycoproteins. At neutral and alkaline pH, Con A exists as a tetramer of four identical subunits; below pH 5.6, Con A dissociates into active dimers of 52 kDa. Acetylation, succinylation, or other derivatizations can also produce stable forms with dimeric structures. (See succinylated Con A). Nicks in the sequence are often present in the purest preparations due to hydrolytic damage within the seeds.
Con A requires calcium or manganese ions at each of its four saccharide binding sites. Although these divalent metal ions are bound tightly to the polypeptide structure, buffers which can bind calcium (such as phosphate) generally should be avoided in diluting Con A, since a gradual loss in activity may occur.
Inhibiting/Eluting Sugar: mixture of 200 mM α-methylmannoside/200 mM α-methylglucoside