[KO验证抗体] Rabbit Anti-CK18 antibody

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K1C18_HUMAN; Keratin, type I cytoskeletal 18; KRT18; CYK18; Cell proliferation-inducing gene 46 protein; Cytokeratin-18 (CK-18); Keratin-18 (K18); PIG46; Keratin 18;

Cat:

SL1339R

Species Reactivity：

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

Immunogen：

KLH conjugated synthetic peptide derived from human CK18:301-423/430

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=0.2μg /testICC=1:100IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Lovo (mouse) Lysate at 40 ugPrimary: Anti- CK18(SL1339R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size:48kDObserved band size: 47kDSample: placenta (Mouse) Lysate at 40 ugPrimary: Anti-CK18 (Bs-1339R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 48 kDObserved band size: 48 kDSample: Kidney (mouse) Lysate at 40 ugPrimary: Anti- CK18(SL1339R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size:48kDObserved band size: 47kDSample: Intestinal (Mouse) Lysate at 30 ugPrimary: Anti- CK18 (SL1339R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilutionPredicted band size: 48 kDObserved band size: 48 kDSample: Lane 1: Hela (Human) Cell Lysate at 30 ugLane 2: HepG2 (Human) Cell Lysate at 30 ugLane 3: A549 (Human) Cell Lysate at 30 ugLane 4: MCF-7 (Human) Cell Lysate at 30 ugPrimary: Anti-CK18 (SL1339R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 48 kDObserved band size: 45 kDSample: Hela KO CK18 (Human) Cell Lysate at 30 ugHela(Human) Cell Lysate at 30 ugPrimary: Anti-CK18 (SL1339R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 48 kDObserved band size: 48 kDParaformaldehyde-fixed, paraffin embedded (human pancreatic cancer); 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 (CK18) Polyclonal Antibody, Unconjugated (SL1339R) 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 uterus); 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 (CK18) Polyclonal Antibody, Unconjugated (SL1339R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.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,SLC0005) at 37℃ for 20 min; Incubation: Anti-CK18 Polyclonal Antibody, Unconjugated(SL1339R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingTissue/cell:A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,SLC0005) at 37°C for 20 min; Antibody incubation with (CK18) polyclonal Antibody, Unconjugated (SL1339R) 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, C02-04002) was used to stain the cell nuclei.Tissue/cell:A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,SLC0005) at 37°C for 20 min; Antibody incubation with (CK18) polyclonal Antibody, Unconjugated (SL1339R) 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, C02-04002) was used to stain the cell nuclei.Blank control (blue line): Hep G2 (blue). Primary Antibody (green line): Rabbit Anti-CK18 antibody (SL1339R) Dilution: 0.2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% methanol for 20 min at -20℃. 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. Acquisition of 20,000 events was performed.Blank control:Hela. Primary Antibody (green line): Rabbit Anti-CK18 antibody (SL1339R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF647Dilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST 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.

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Unit:

Price: $236.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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KRT18 encodes the type I intermediate filament chain keratin 18. Keratin 18, together with its filament partner keratin 8, are perhaps the most commonly found members of the intermediate filament gene family. They are expressed in single layer epithelial tissues of the body. Mutations in this gene have been linked to cryptogenic cirrhosis. Two transcript variants encoding the same protein have been found for this gene. [provided by RefSeq, Jul 2008]

Function:
When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection.

Post-translational modifications:
Phosphorylation increases by IL-6.
Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-231 by either caspase-3, caspas-6 or caspase-7.
O-glycosylated at multiple sites; glycans consist of single N-acetylglucosamine residue.

Similarity:
Belongs to the intermediate filament family.

SWISS:
P05783

Gene ID:
3875

Database links:

Entrez Gene: 50696 Cow

Entrez Gene: 3875 Human

Entrez Gene: 16668 Mouse

Entrez Gene: 294853 Rat

SwissProt: P05783 Human

SwissProt: P05784 Mouse

SwissProt: Q5BJY9 Rat

结构蛋白（Structural Proteins）
常用于肿瘤细胞的分化、增殖及转移方面的研究。 Picture

Sample: Lovo (mouse) Lysate at 40 ug
Primary: Anti- CK18(SL1339R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size:48kD
Observed band size: 47kD

Sample:
placenta (Mouse) Lysate at 40 ug
Primary: Anti-CK18 (Bs-1339R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 48 kD

Sample: Kidney (mouse) Lysate at 40 ug
Primary: Anti- CK18(SL1339R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size:48kD
Observed band size: 47kD

Sample: Intestinal (Mouse) Lysate at 30 ug
Primary: Anti- CK18 (SL1339R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilution
Predicted band size: 48 kD
Observed band size: 48 kD

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 (SL1339R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 45 kD

Sample:
Hela KO CK18 (Human) Cell Lysate at 30 ug
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-CK18 (SL1339R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 48 kD

Paraformaldehyde-fixed, paraffin embedded (human pancreatic cancer); 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 (CK18) Polyclonal Antibody, Unconjugated (SL1339R) 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 uterus); 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 (CK18) Polyclonal Antibody, Unconjugated (SL1339R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

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,SLC0005) at 37℃ for 20 min;
Incubation: Anti-CK18 Polyclonal Antibody, Unconjugated(SL1339R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

Tissue/cell:A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,SLC0005) at 37°C for 20 min; Antibody incubation with (CK18) polyclonal Antibody, Unconjugated (SL1339R) 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, C02-04002) was used to stain the cell nuclei.

Blank control (blue line): Hep G2 (blue).
Primary Antibody (green line): Rabbit Anti-CK18 antibody (SL1339R)
Dilution: 0.2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% methanol for 20 min at -20℃. 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. Acquisition of 20,000 events was performed.

Blank control:Hela.
Primary Antibody (green line): Rabbit Anti-CK18 antibody (SL1339R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST 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.

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