Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-PPAR alpha antibody (orb15031), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
Blank control: Jurkat. Primary Antibody (green line): Rabbit Anti-PPAR alpha antibody (orb15031), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
Paraformaldehyde-fixed, paraffin embedded (rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (PPAR alpha) Polyclonal Antibody, Unconjugated (orb15031) at 1:200 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Rat splenocytes stained with Anti-PPAR alpha Polyclonal Antibody, PE-CY5 Conjugated at 1:50.
Sample: Heart (Mouse) Lysate at 40 ug Heart (Rat) Lysate at 40 ug Liver (Mouse) Lysate at 40 ug Urinary bladder (Mouse) Lysate at 40 ug Kidney (Mouse) Lysate at 40 ug Primary: Anti-PPAR alpha (orb15031) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 52/19 kD Observed band size: 52 kD.
Sample: Heart (Mouse) Lysate at 40 ug, Primary: Anti-PPAP alpha (orb15031) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution, Predicted band size: 51 kD, Observed band size: 51 kD.
Sample: Lane 1: Heart (Mouse) Lysate at 30 ug, Lane 2: Liver (Mouse) Cell Lysate at 30 ug, Primary: Anti-PPAR alpha (orb15031) at 1:300 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1:5000dilution, Predicted band size: 51 kD, Observed band size: 51 kD.
Flow cytometric analysis of rat splenocytes (Dilution at:1:50) using PPAR alpha antibody
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