Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human,Rat (predicted:Mouse,Chicken,Dog,Rabbit)

IgG

Human,Rat (predicted:Mouse,Chicken,Dog,Rabbit)

1. Sample:
Hela KO CK18 (Human) Cell Lysate at 30 μg
Hela (Human) Cell Lysate at 30 μg
Primary: Anti-CK18 (TMAB-01037) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 48 kDa
2. Sample:
Lane 1: Hela (Human) Cell Lysate at 30 μg
Lane 2: HepG2 (Human) Cell Lysate at 30 μg
Lane 3: A549 (Human) Cell Lysate at 30 μg
Lane 4: MCF-7 (Human) Cell Lysate at 30 μg
Primary: Anti-CK18 (TMAB-01037) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 45 kDa
3. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-CK18 antibody (TMAB-01037)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at-20°C. 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.
4. Paraformaldehyde-fixed, paraffin embedded Human Pancreatic Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CK18 Polyclonal Antibody, Unconjugated (TMAB-01037) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Rat Uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CK18 Polyclonal Antibody, Unconjugated (TMAB-01037) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (CK18) polyclonal Antibody, Unconjugated (TMAB-01037) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
7. Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CK18 Polyclonal Antibody, Unconjugated (TMAB-01037) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
8. Blank control (blue line): HepG2 (blue).
Primary Antibody (green line): Rabbit Anti-CK18 antibody (TMAB-01037)
Dilution: 0.2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% methanol for 20 min at-20°C. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 0.2μg /test

Polyclonal

KLH conjugated synthetic peptide: human CK18

Human

3875

Cytokeratins,Tumor Associated