Flow cytometric analysis of Hepg2 Cells using Cytochrome C antibody.
Immunohistochemical staining of rat brain tissue using Cytochrome C antibody.
Blank control: HepG2 (blue). Primary Antibody: Rabbit Anti-Cytochrome C antibody (orb221350, Green), dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue), dilution: 1:200 in 1X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde for 10 min at 37C. Primary antibody (orb221350, 1 µg/1x10 6 cells) were incubated for 30 min at room temperature, followed by 1X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature. Acquisition of 20000 events was performed.
Sample: Cytochrome C protein at 30 ug, Primary: Anti-Cytochrome C (orb221350) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 14 kD, Observed band size: 14 kD.
Sample: Lane 1: Heart (Rat) Lysate at 40 ug, Lane 2: Heart (Mouse) Lysate at 40 ug, Primary: Anti-Cytochrome C (orb221350) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 14.4 kD, Observed band size: 14.4 kD.
Tissue/Cell: Sh-sy5y cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (Cytochrome C) polyclonal Antibody, Unconjugated (orb221350) 1:100, 90 minutes at 37C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
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