WB
Western blot analysis of MOG on different lysates with Rabbit anti-MOG antibody at 1/1,000 dilution.
Lane 1: Mouse brain tissue lysate (40 µg/Lane)
Lane 2: Mouse lung tissue lysate (negative) (40 µg/Lane)
Lane 3: Rat brain tissue lysate (40 µg/Lane)
Lane 4: Rat lung tissue lysate (negative) (40 µg/Lane)
Exposure time: 1 minute;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-MOG antibody at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IF-F
Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling MOG with Rabbit anti-MOG antibody.
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/50 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
IF-P
Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling MOG with Rabbit anti-MOG antibody at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/200 dilution overnight at 4 ℃, washed with PBS.
Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).