All lanes : Anti-PPAR alpha (phospho S12) antibody (ab3484) at 1/200 dilution

Lane 1 : C2C12 cell lysate
Lane 2 : NIH-3T3 cell lysate
Lane 3 : 3T3-L1 cell lysate
Lane 4 : HepG2 cell lysate

Lysates/proteins at 25 µg per lane.

Predicted band size: 52 kDa
Observed band size: 52 kDa

Immunofluorescent analysis of Phospho-PPAR alpha pSer12 (green) showing staining in the nucleus of C2C12 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-PPAR alpha pSer12 polyclonal antibody (ab3484) 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-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Immunofluorescent analysis of Phospho-PPAR alpha pSer12 (green) showing staining in the nucleus of 3T3-L1 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-PPAR alpha pSer12 polyclonal antibody (ab3484) 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-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

All lanes : Anti-PPAR alpha (phospho S12) antibody (ab3484) at 1/1000 dilution

Lane 1 : U-87 MG with Skimmed milk
Lane 2 : MCF7 with Skimmed milk
Lane 3 : MDA-MB-231 with Skimmed milk
Lane 4 : C2C12 with Skimmed milk
Lane 5 : Hep G2 with Skimmed milk
Lane 6 : NIH/3T3 with Skimmed milk

Lysates/proteins at 20 µg per lane.

Blocking peptides at 5 % per lane.

Secondary
All lanes : Goat anti-rabbit IgG (H+L) at 1/2500 dilution

Predicted band size: 52 kDa

Immunofluorescent analysis of Phospho-PPAR alpha pSer12 (green) showing staining in the nucleus of U-87 MG cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-PPAR alpha pSer12 polyclonal antibody (ab3484) 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-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

ab3484 staining PPAR alpha (phospho S12) in Mouse neuronal cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. A Cy2^®-conjugated Donkey anti-rabbit IgG polyclonal (1/100) was used as the secondary antibody.

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ab3484 staining PPAR alpha (phospho S12) in MCF7 cells by Flow Cytometry. The sample was incubated with the primary antibody (3-5 ug/million cells in 2.5% BSA) for 2 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit was used as the secondary antibody (1/400). Red histogram represents ab3484, pink histogram represents isotype control, purple histogram represents unstained control and green histogram represents no primary antibody control.