PPAR gamma Antibody

Immunofluorescent analysis of PPAR gamma (green) showing positive staining in the nucleus and cytoplasm of 3T3-L1 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with a PPAR gamma polyclonal antibody (Product # PA3-821A) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-rabbit IgG secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Immunofluorescent analysis of PPAR gamma (green) showing positive staining in the nucleus and cytoplasm of Hela cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with a PPAR gamma polyclonal antibody (Product # PA3-821A) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-rabbit IgG secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Immunofluorescent analysis of PPAR gamma (green) showing positive staining in the nucleus and cytoplasm of NIH-3T3 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with a PPAR gamma polyclonal antibody (Product # PA3-821A) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-rabbit IgG secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Western blot analysis of PPAR was performed by loading 25 ug of 3T3-L1 (Lane 1), NIH-3T3 (Lane 2), and Hela (Lane 3) cell lysates and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4ºC overnight. The membrane was probed with a PPAR polyclonal antibody (Product # PA3-821A) at a dilution of 1:1000 (3T3-L1 and Hela) and 1:500 (NIH-3T3) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Results show a band at 57 kDa in all three cell lysates.