WB
Western blot analysis of TGF beta 1 on different lysates with Rabbit anti-TGF beta 1 antibody at 1/1,000 dilution.
Lane 1: Raji cell lysate,
Lane 2: A549 cell lysate,
Lane 3: L-929 cell lysate,
Lane 4: NIH/3T3 cell lysate,
Lane 5: Mouse spleen tissue lysate,
Lysates/proteins at 20 µg/Lane.
Exposure time: 2 minutes 50 seconds;
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 spleen tissue with Rabbit anti-TGF beta 1 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.