[IF=2.906] Jinfeng Zhu et al. Expression and prognostic significance of pyruvate dehydrogenase kinase 1 in bladder urothelial carcinoma. Virchows Arch. 2020 Nov;477(5):637-649.  IHC ;  Human.  

PubMed:32388719

[IF=2.705] Hu HJ et al. Regulating the Warburg effect on metabolic stress and myocardial fibrosis remodeling and atrial intracardiac waveform activity induced by atrial fibrillation. Biochem Biophys Res Commun. 2019 Aug 27;516(3):653-660.  WB ;  beagle canine.  

PubMed:31242971

Sample:
Lane 1: Mouse Heart tissue lysates
Lane 2: Rat Heart tissue lysates
Primary: Anti-PDK1/Mitochondrial Pyruvate dehydrogenase kinase 1 (bs-3788R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 50 kDa

Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PDK1/Mitochondrial Pyruvate dehydrogenase kinase 1) Polyclonal Antibody, Unconjugated (bs-3788R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control（black line）:SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-PDK1 antibody (bs-3788R)
Dilution:1ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Isotype control（orange line）: Normal Rabbit IgG
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 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.