Sample:
Lung (Mouse) Lysate at 40 ug
Primary: Anti-HIF3 alpha (SL5898R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 74 kD
Observed band size: 75 kD
Paraformaldehyde-fixed, paraffin embedded (Rat kidney); 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 (HIF3 alpha) Polyclonal Antibody, Unconjugated (SL5898R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control (blue line): A549 (fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-HIF3 alpha antibody (SL5898R),ilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE,Dilution: 1μg /test.
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