|
A549 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 (Cytokeratin 7) polyclonal Antibody, Unconjugated (orb13342) 1:100, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei. |
|
Blank control: Hela. Primary Antibody (green line): Rabbit Anti-Cytokeratin 7 antibody (orb13342), Dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. 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 20000 events was performed. |
|
Paraformaldehyde-fixed, paraffin embedded (Rat bladder), 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 (CK7) Polyclonal Antibody, Unconjugated (orb13342) at 1:200 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
|
Paraformaldehyde-fixed, paraffin embedded (Rat placenta), 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 (CK7) Polyclonal Antibody, Unconjugated (orb13342) at 1:200 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
|
Sample: Lane 1: Mouse Urinary bladder tissue lysates, Lane 2: Mouse Breast tissue lysates, Lane 3: Mouse Placenta tissue lysates, Lane 4: Mouse trachea tissue lysates, Lane 5: Rat Urinary bladder tissue lysates, Lane 6: Rat Placenta tissue lysates, Lane 7: Human Hela cell lysates, Lane 8: Human HepG2 cell lysates, Lane 9: Human Siha cell lysates, Lane 10: Human Huvec cell lysates, Primary: Anti-CK7 (orb13342) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 54 kDa, Observed band size: 52 kDa. |
|
Tissue/Cell: human lung carcinoma, 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-Cytokeratin 7 Polyclonal Antibody, Unconjugated (orb13342) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |