Analysis of complement factor B activation by PAGE-immunoblotting.
Initiation of complement activation via the alternative pathway results in cleavage of Factor B into two fragments Ba and Bb. Employing PAGE and immunoblotting analysis we have demonstrated Ba and Bb activation fragments following in vitro and in vivo activation in a range of biological fluids including cerebrospinal fluid and synovial fluid. Quantitative linearity was obtained over the range 5-20 micrograms/ml of Factor B and its fragments in body fluids. Using Davis-PAGE a previously unrecognised triple component heterogeneity of the Ba fragment was observed. This study demonstrates that immunoblotting may be used as a sensitive and quantitative tool for the detection and characterisation of the individual cleavage products of Factor B in complex biological fluids without prior purification.
Doran
M
M
Children's Research Centre, Our Lady's Hospital, Dublin, Ireland.
Reen
D J
DJ
D002315:Cardiopulmonary Bypass; D003167:Complement Activation; D011415:Complement Factor B / Q000032:analysis / Q000378:metabolism*; D003720:Densitometry; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004792:Enzyme Precursors / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D012967:Sodium Dodecyl Sulfate
D004792:Enzyme Precursors; D012967:Sodium Dodecyl Sulfate; D011415:Complement Factor B