WB
Western blot analysis of Phospho-Glycogen synthase 1 (S641) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: SK-Br-3 cell lysate,
Lane 2: A431 cell lysate.
WB
Western blot analysis of Phospho-Glycogen synthase 1 (S641) on HepG2 cell lysates.
Lane 1: HepG2 cells, whole cell lysate, 10ug/lane,
Lane 2: HepG2 cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane,
All lanes :
Anti-Phospho-Glycogen synthase 1 (S641) antibody at 1/500 dilution. Anti-GAPDH antibody at 1/10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) at 1/200,000 dilution.
Blocking and diluting buffer: 5% BSA.
Exposure time: 1 minute 34 seconds.
WB
Western blot analysis of Phospho-Glycogen synthase 1 (S641) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Mouse liver tissue lysate, untreated,
Lane 2: Mouse liver tissue lysate, treated with AP.
ICC/IF
ICC staining of Phospho-Glycogen synthase 1 (S641) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC/IF
ICC staining of Phospho-Glycogen synthase 1 (S641) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Phospho-Glycogen synthase 1 (S641) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-Phospho-Glycogen synthase 1 (S641) 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.