The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA: Use at a concentration of 0.5 - 1.0 µg/ml. The detection limit for recombinant human IGFBP4 is approximately 0.06 ng/well.
Neut: Use at an assay dependent dilution.
WB: Use at a concentration of 0.1 - 0.2 µg/ml. Detects bands of approximately 25 and 32 kDa. The detection limit for recombinant human IGFBP4 is approximately 2 ng/lane and 5 ng/lane under non-reducing and reducing conditions, respectively.
The Neutralization Dose50 (ND50) for anti human IGFBP4 is approximately 10 to 30 µg/ml in the presence of 0.3 µg/ml of recombinant human IGFBP4 and 14 ng/ml of recombinant human IGF-II using the human breast cancer cell line MCF-7 cells. The ND50 is the concentration of antibody required to yield one half maximal inhibition of the cytokine activity on a responsive cell line, when the cytokine is present at a concentration just high enough to elicit a maximum response. The exact concentration of antibody required to neutralize human IGFBP4 activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity studied.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
IGF-binding proteins prolong the half-life of the IGFs and have been shown to either inhibit or stimulate the growth promoting effects of the IGFs on cell culture. They alter the interaction of IGFs with their cell surface receptors.