The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2 µg/ml. Predicted molecular weight: 37 kDa.
Use a concentration of 5 µg/ml. (paraformaldehyde fixed cells)
Western blot - Anti-AKR1C1 antibody [4B6AF3] (ab131375)
All lanes : Anti-AKR1C1/AKR1C2 antibody [4B6AF3] (ab131375) at 2 µg/ml
Lane 1 : Molecular weight marker Lane 2 : HeLa Cell Lysate at 15 µg Lane 3 : HepG2 Cell Lysate at 15 µg Lane 4 : Human Liver Homogenate (HLH) at 10 µg Lane 5 : Human Heart Homogenate (HHH) at 10 µg Lane 6 : Mouse Liver Homogenate (MLH) at 10 µg Lane 7 : Rat Liver Homogenate (RLH) at 10 µg Lane 8 : Rat Heart Homogenate (RHH) at 10 µg
Immunofluorescence analysis of AKR1C1 in HeLa cells, using ab131375 at 5 µg/ml. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 min). The cells were then incubated with ab131375 for 2h at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1h. 1% BSA was used as the blocking agent for all blocking steps. The target protein locates to the cytosol.