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[IF=2.24] Zhang, Ni-Ni, et al. "Functional regeneration of irradiated salivary glands with human amniotic epithelial cells transplantation." International Journal of Clinical and Experimental Pathology? 6.10 (2013): 2039-2047.  Mouse.  

PubMed:24133581

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 (bs-1744R) 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 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-Cytokeratin 7 Polyclonal Antibody, Unconjugated(bs-1744R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Paraformaldehyde-fixed, paraffin embedded (Rat bladder); 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 (CK7) Polyclonal Antibody, Unconjugated (bs-1744R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat placenta); 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 (CK7) Polyclonal Antibody, Unconjugated (bs-1744R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Cytokeratin 7) polyclonal Antibody, Unconjugated (bs-1744R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control:Hela. Primary Antibody (green line): Rabbit Anti-Cytokeratin 7 antibody (bs-1744R) Dilution: 1ug/Test; Secondary Antibody : Goat anti-rabbit IgG-FITC Dilution: 0.5ug/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.