The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
EIA: Use at an assay dependent dilution.
ICC/IF: Use at a concentration of 1 µg/ml.
IP: Use at an assay dependent dilution.
RIA: Use at an assay dependent dilution.
WB: Use at an assay dependent dilution. Predicted molecular weight: 46 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
ICC/IF image of ab48646 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab48646, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Thyroxine Binding Globulin antibody (ab48646)
Anti-Thyroxine Binding Globulin antibody (ab48646) at 1 µg/ml + Natural Human Thyroxine Binding Globulin protein (ab96516) at 0.01 µg