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References in periodicals archive ?
We added 100 [micro]L of mouse monoclonal anti-H-ficolin (clone 4H5, HyCult Biotechnology b.v.), rabbit polyclonal anti-L-ficolin (Tokai University), or DIG-labeled anti-MBL, at 500-fold dilution, in blocking buffer.
After blocking the endogenous peroxidase with 30 g/L hydrogen peroxide, for ficolin staining alone, sections were incubated in TBS with normal sheep serum, with either H-ficolin monoclonal antibody (clone 4H5) from HyCult Biotechnology or rabbit polyclonal anti-L-ficolin antibody (clone 2F5) from Tokai University.
To test the specificity and cross-reactivity of the antibodies used in the ELISA, we subjected samples containing B- and H-FABP (both 1 [micro]g and 10 [micro]g in each case) to 13.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotted the gels on a HyBond nitrocellulose membrane (Amersham Biosciences), and detected the FABPs with purified polyclonal antibodies against B-FABP and with monoclonal antibodies against H-FABP (67D3 and 66E2 as capture and detector antibodies, respectively, in the H-FABP ELISA; HyCult Biotechnology) (23,24).
Tissue and plasma concentrations of H-FABP were measured with a sandwich-type ELISA obtained from HyCult Biotechnology (HK 403) exactly as described previously (21, 23).
Briefly, the procedure involved coating a microtiter ELISA plate with monoclonal detector antibody 67D3 (gift from HyCult Biotechnology, Uden, The Netherlands) and incubating for 1 h with recombinant calibrator or perfusate before the addition of a second, horseradish peroxidase-labeled detector antibody, 66E2-HRP (HyCult Biotechnology).
L-FABP was measured with a sandwich ELISA developed in collaboration with HyCult Biotechnology (Uden, The Netherlands).