[IF=1.772] Penggang Liu. et al. Expression of tumour transcription factor GLI1 in canine mammary tumours tissue. VET MED SCI. 2022 Jun 06  WB ;  Dog.  

PubMed:35667035

Sample:
Lane 1: Mouse NIH/3T3 cell lysates
Lane 2: Human MCF-7 cell lysates
Primary: Anti-PCGF4/BMI1 (bs-2999R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 44 kDa

Paraformaldehyde-fixed, paraffin embedded (Human lung cancer); 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 (BMI1 ) Polyclonal Antibody, Unconjugated (bs-2999R) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody (sp-0023) for 20 minutes and DAB staining.

Blank control（black line）:NIH/3T3.
Primary Antibody (green line): Rabbit Anti-PCGF4/BMI1 antibody (bs-2999R)
Dilution:1ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488
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.

Blank control（black line）:NIH/3T3.
Primary Antibody (green line): Rabbit Anti-PCGF4/BMI1 antibody (bs-2999R)
Dilution:1ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488
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.