Phospho-RPS6 (S235 + S236) Recombinant Rabbit Monoclonal Antibody [PS01-04]

Western blot analysis of Phospho-RPS6 (S235 + S236) on different lysates with Rabbit anti-Phospho-RPS6 (S235 + S236) antibody (HA721275) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate
Lane 2: THP-1 cell lysate
Lane 3: Hela cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 29 kDa
Observed band size: 33 kDa

Exposure time: 30 seconds;

12% 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 (HA721275) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-RPS6 (S235 + S236) antibody (HA721275) 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 (HA721275) 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.

Immunocytochemistry analysis of Hela cells labeling Phospho-RPS6 (S235 + S236) with Rabbit anti-Phospho-RPS6 (S235 + S236) antibody (HA721275) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-RPS6 (S235 + S236) antibody (HA721275) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.