2010
DOI: 10.1080/15321811003617305
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Elisa for Rankl-Opg Complex in Mouse Sera

Abstract: A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) for the mouse RANKL-OPG complex was developed by utilizing a polyclonal antibody that recognizes mouse soluble RANKL as an immobilized capture component and mouse OPG IgG labeled with peroxidase. We could quantify the RANKL-OPG complex level (detection limit: 1 pmol/L). Employing this assay system, we demonstrated that the RANKL-OPG complex was constitutively present in the serum of OPG+/- mice, but not in that of OPG-/- or wild-type C57BL/6 J mice. Show more

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Cited by 4 publications
(4 citation statements)
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“…Complexes of RANKL-OPG were analyzed in a modified procedure, using reagents from the same ELISAs (i.e., anti-RANKL capture antibodies and anti-OPG detection antibodies). The procedure has been previously described in detail ( Mogi and Kondo, 2010 ). Optical densities were measured using a BioTek Synergy HT (BioTek Instruments, Vermont, USA), and wavelength correction was applied by subtracting readings at 540 nm from those at 450 nm.…”
Section: Methodsmentioning
confidence: 99%
“…Complexes of RANKL-OPG were analyzed in a modified procedure, using reagents from the same ELISAs (i.e., anti-RANKL capture antibodies and anti-OPG detection antibodies). The procedure has been previously described in detail ( Mogi and Kondo, 2010 ). Optical densities were measured using a BioTek Synergy HT (BioTek Instruments, Vermont, USA), and wavelength correction was applied by subtracting readings at 540 nm from those at 450 nm.…”
Section: Methodsmentioning
confidence: 99%
“…Secreted IL-6 (DY205), CXCL8 (DY208), DCN (DY143), TGF-β1 (DY240), Osteoprotegerin (OPG; DY805), receptor activator of nuclear factor kappa-B ligand (RANKL; DY626) and Connective Tissue Growth Factor (CTGF) (DY9190-05) were measured using Human DuoSet Enzyme linked immunosorbent assay (ELISA) kits (R&D Systems, Abingdon, United Kingdom) according to the manufacturer's instructions. Total RANKL was calculated by measuring free RANKL and in complex with OPG using an in house developed OPG/RANKL complex sandwich ELISA by using a human capture RANKL antibody (part of cat#DY626, R&D Systems) and a human detection OPG antibody (part of cat#DY805, R&D Systems) (Mogi and Kondo, 2010). Total OPG was calculated by measuring free OPG and bound OPG.…”
Section: Enzyme Linked Immunosorbent Assaymentioning
confidence: 99%
“…[18][19][20] Briefly, 100 mL=well of the capture anti-human TRAIL monoclonal IgG (2 mg=mL of 50 mM carbohydrate buffer, pH 9.6) was transferred to an ELISA plate (Nunc, Denmark, 96 wells, flat bottom) and incubated at 37 C for 1 hr; then each well was aspirated, and the plate was blocked by adding 150 mL phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA, buffer-1) to each well. Incubation was carried out at 37 C for a minimum of 1 hr.…”
Section: Elisa Proceduresmentioning
confidence: 98%