[IF=4.932] Yi Ren. et al. Pravastatin attenuates sepsis-induced acute lung injury through decreasing pulmonary microvascular permeability via inhibition of Cav-1/eNOS pathway. Int Immunopharmacol. 2021 Nov;100:108077  WB ;  Mouse.  

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[IF=4.599] Juan Tan. et al. Circ_0124644 Serves as a ceRNA for miR-590-3p to Promote Hypoxia-Induced Cardiomyocytes Injury via Regulating SOX4. Front Genet. 2021; 12: 667724  WB ;  Human.  

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[IF=2.523] Zhi Chen. et al. Klotho deficiency aggravates diabetes-induced podocyte injury due to DNA damage caused by mitochondrial dysfunction. Int J Med Sci. 2020; 17(17): 2763–2772  WB ;  Mouse.  

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Sample:
Lung (Mouse) Lysate at 40 ug
Primary: Anti- caspase-3 p17 subunit (bs-20364R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17/32 kD
Observed band size: 17/32 kD

Sample:
Spleen (Mouse) Lysate at 40 ug
NIH/3T3 (Mouse) Cell Lysate at 30 ug
Heart (Mouse) Lysate at 40 ug
Primary: Anti- caspase-3 p17 subunit (bs-20364R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17/32 kD
Observed band size: 17/32 kD

Sample:
Jurkat (human) Cell Lysate at 30 ug
Primary: Anti- caspase-3 p17 subunit (bs-20364R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17/32 kD
Observed band size: 17/32 kD

Paraformaldehyde-fixed, paraffin embedded (Human glioma); 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 (caspase-3 p17 subunit) Polyclonal Antibody, Unconjugated (bs-20364R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control (blue line): Hela (fixed with 80% methanol (5 min at -20℃) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-caspase-3 p12 subunit antibody (bs-20364R),Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC,Dilution: 1μg /test.

Overlay histogram showing HL 60 cells stained with bs-20364R (Green line).
The cells were fixed with 90% methanol (5 min) and then permeabilized with 0.01M PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (bs-20364R,1μg/1x10^6 cells) for 30 min at 22℃. The secondary antibody used was fluorescein isothiocyanate goat anti-rabbit IgG (H+L) (bs- 0295G-FITC , Brillant blue line) at 1/200 dilution for 30 min at 22℃. Isotype control antibody was rabbit IgG (polyclonal,bs-0295P,Orange line) (1μg/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of 20,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Blank control: NIH/3T3.
Primary Antibody (green line): Rabbit Anti-caspase-3 p17 subunit antibody (bs-20364R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
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 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.