[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:
Placenta (Mouse) Lysate at 30 ug
Lung (Mouse) Lysate at 30 ug
Primary: Anti-Cytokeratin 7 (bs-1744R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilution
Predicted band size: 54 kD
Observed band size: 54 kD

Sample: Large intestine (Mouse) Lysate at 30 ug
Primary: Anti- Cytokeratin 7 (bs-1744R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilution
Predicted band size: 54 kD
Observed band size: 54 kD

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

Sample:
Large intestine (Mouse) Lysate at 40 ug
Large intestine(Rat) Lysate at 40 ug
Primary: Anti- Cytokeratin 7 (bs-1744R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 54 kD
Observed band size: 54 kD

Sample:
Embryo (Mouse) Lysate at 40 ug
Primary: Anti- Cytokeratin 7 (bs-1744R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 54 kD
Observed band size: 54 kD

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.