Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human,Mouse,Rat (predicted:Chicken,Pig,Cow,Rabbit,Sheep)

IgG

Human,Mouse,Rat (predicted:Chicken,Pig,Cow,Rabbit,Sheep)

1. Sample:
Lane 1: Mouse Breast tissue lysates
Lane 2: Mouse Lung tissue lysates
Lane 3: Rat Adipose tissue lysates
Lane 4: Rat Breast tissue lysates
Lane 5: Rat Lung tissue lysates
Lane 6: Human A431 cell lysates
Lane 7: Human A549 cell lysates
Lane 8: Human THP-1 cell lysates
Lane 9: Human 293T cell lysates
Primary: Anti-PPAR gamma (TMAB-01568) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 52 kDa
2. Tissue/cell: mouse lung tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-PPAR Gamma Polyclonal Antibody, Unconjugated (TMAB-01568) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Blank control (blue line): U251
Primary Antibody (green line): Rabbit Anti-PPARG/PPAR gamma antibody (TMAB-01568)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
4. Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (PPAR gamma) Polyclonal Antibody, Unconjugated (TMAB-01568) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Blank control: A431. Primary Antibody (green line): Rabbit Anti-PPAR gamma antibody (TMAB-01568)
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-20°C. 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.
6. Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (PPAR gamma) polyclonal Antibody, Unconjugated (TMAB-01568) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human PPAR Gamma

Human

5468

Diabetes associated,Metabolism,Zinc Finger,Fatty acids,Fatty acid oxidation,Diabetes,Heart disease,Obesity,Metabolism,Adipogenesis