CK18 Rabbit Polyclonal Antibody

• Catalog Number: orb500815
• Category: Antibodies
• Description: CK18 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Canine, Gallus, Mouse, Rabbit
• Reactivity: Human, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human CK18 (301-423/430aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=0.2μg /test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 48 kDa
• Target: KRT18
• UniProt ID: P05783
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: K1C18_HUMAN; Keratin, type I cytoskeletal 18; KRT1
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Sample: Lane 1: Hela (Human) Cell Lysate at 30 ug, Lane 2: HepG2 (Human) Cell Lysate at 30 ug, Lane 3: A549 (Human) Cell Lysate at 30 ug, Lane 4: MCF-7 (Human) Cell Lysate at 30 ug, Primary: Anti-CK18 (orb500815) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 45 kD.

Blank control (blue line): Hep G2 (blue). Primary Antibody (green line): Rabbit Anti-CK18 antibody (orb500815), 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 1X 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. Acquisition of 20000 events was performed.

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-CK18 antibody (orb500815), 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. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human pancreatic cancer), 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 37°C for 30 min, Antibody incubation with (CK18) Polyclonal Antibody, Unconjugated (orb500815) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat uterus), 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 37°C for 30 min, Antibody incubation with (CK18) Polyclonal Antibody, Unconjugated (orb500815) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Hela KO CK18 (Human) Cell Lysate at 30 ug, Hela (Human) Cell Lysate at 30 ug, Primary: Anti-CK18 (orb500815) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 48 kD.

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 (orb500815) 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 nuclei.