Distinction between classical and alternative pathways

In experimental work, the two pathways of activation can be readily distinguished, in that both pathways require Mg2+, but only the classical pathway requires Ca2+ ions: if these are chelated by EGTA (ethyleneglycol-bis-(|3-amino-ethylether) tetra-acetic acid) and Mg2+ ions are added in excess, only the alternative pathway will function. Further, the activation of the alternative pathway is prevented by dilution of human serum beyond about ten-fold, and is generally considerably impaired after 2-3 days storage of human serum at 4°C: the classical pathway, via Clq, operates at lower concentrations of serum and is stabler on storage. The stability of activation occurring via MBL has not been extensively examined.

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