Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (TMEM176A) Polyclonal Antibody, Unconjugated (orb158621) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Protein: HepG2 (human) lysates at 30 ug, Primary: rabbit Anti-TMEM176A (orb158621) at 1:300, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1:5000, Predicted band size: 26 kD, Observed band size: 26 kD.
Protein: mouse kidney lysates, 30 ug, Primary: Anti-TMEM176A (orb158621) at 1:300, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1:5000, ECL excitated the fluorescence, Predicted band size: 24kD, Observed band size: 24kD.
Sample: U937 (Human) Cell Lysate at 30 ug, Primary: Anti-TMEM176A (orb158621) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 26 kD, Observed band size: 25 kD.
Tissue/Cell: mouse embryo tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-TMEM176A Polyclonal Antibody, Unconjugated (orb158621) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining.
Western blot analysis of HepG2 lysates using TMEM176A antibody
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