[IF=4.398] Xiaoyu Qu. et al. Integration of metabolomics and proteomics analysis to explore the mechanism of neurotoxicity induced by receipt of isoniazid and rifampicin in mice. NEUROTOXICOLOGY. 2023 Jan;94:24  IHC ;  Mouse.  

PubMed:36347327

Sample:
Lane 1: Mouse Placenta tissue lysates
Lane 2: Mouse Testis tissue lysates
Primary: Anti-PPAR delta + beta (bs-0250R) at 1/200 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 53 kDa

Tissue/cell: Human esophageal carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-PPAR-delta Polyclonal Antibody, Unconjugated(bs-0250R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control: A431.
Primary Antibody (green line): Rabbit Anti-PPAR delta + beta antibody (bs-0250R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at 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 20,000 events was performed.