Rabbit Monoclonal Cytokeratin 18 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 35 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/2000 | Notes For unpurified use at 1/10000 Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20 | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection.
CYK18, PIG46, CYK18, PIG46, KRT18, Cell proliferation-inducing gene 46 protein, Cytokeratin-18, Keratin-18, CK-18, K18
Rabbit Monoclonal Cytokeratin 18 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 35 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
E431-1
Affinity purification Protein A
Human Cytokeratin 18 (K18) was used as immunogen after isolation from cells pre-treated with okadaic acid or pervanadate to promote Tyr hyperphosphorylation.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
Cytokeratin 18 (CK18) also known as Keratin 18 or KRT18 is a type I intermediate filament protein with a molecular weight of approximately 45 kDa. It plays a mechanical role in stabilizing epithelial cell structure and function. CK18 is expressed in single-layered epithelial tissues such as the liver intestine and pancreas. It partners with the type II keratin usually KRT8 to form a stable cytoskeletal network in these cells providing structural integrity and resilience.
CK18 maintains cellular integrity and mediates resistance to mechanical and non-mechanical stresses in epithelial cells. It forms heterodimers with KRT8 participating in a complex network of filaments important for maintaining cell shape and stability. This organization plays a role in managing cell survival during stress and apoptosis helping regulate cellular functions and responses in tissues where it is expressed.
Cytokeratin 18 is involved in the apoptosis and cellular stress response pathways. It interacts with proteins such as caspases during apoptosis facilitating cytoskeletal reorganization and cell fragmentation. CK18 in coordination with KRT8 also participates in signaling pathways that regulate cell cycle and apoptotic processes underlining its importance in cellular turnover and tissue homeostasis.
Elevated levels of CK18 fragments are often observed in cases of non-alcoholic steatohepatitis (NASH) and breast cancer. It serves as a biomarker for liver injury due to its abundance in hepatocytes and release upon cell death. In breast cancer alterations in CK18 expression can reflect disease progression where it interacts with other proteins such as vimentin indicating epithelial-to-mesenchymal transition a process that facilitates metastasis.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Terms & Conditions.
All lanes: Western blot - Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/2000 dilution
Lane 1: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg
Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 48 kDa
Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Cytokeratin 18 with purified ab32118 at 1/500. Cells were fixed with 100% Methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution was used as the secondary antibody. The cells were co-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000) using Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120).
For negative control 2, mouse primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) were used.
Alexa Fluor® 488 (Alexa Fluor® 488 Anti-Cytokeratin 18 antibody [E431-1] ab194124) conjugated version is available for this clone.
Immunocytochemistry/Immunofluorescence analysis of HeLa (+ve) and U87-MG (-ve) cells labelling Cytokeratin 18 with ab32118 at 2 ug/ml overnight at +4°C. Cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. A preadsorbed Alexa Fluor® 488-conjugated goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution was used as the secondary antibody. The cells were co-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), using Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, a preadsorbed Alexa Fluor® 647-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach tissue sections labeling Cytokeratin 18 with purified ab32118 at 1/500 dilution (0.08 µg/ml). Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Indirect immunofluorescence staining with antibodies against DNA, cytokeratin-7 and cytokeratin-18 (upper panel) or against DNA, vimentin and cytokeratin-18 (lower panel). Representatives are presented. Scale bar: 20 μm.
Control or treated cells were fixed for 15 min with 4% PFA containing 0.1% Triton X-100 at room temperature. The following primary antibodies were used for staining: monoclonal mouse antibodies against vimentin and cytokeratin-7 (both 1:100, DAKO) and monoclonal rabbit antibody against cytokeratin-18 (1:50, Abcam). DNA was stained using DAPI (4',6-diamidino-2-phenylindole-dihydrochlorid) (Roche). Slides were examined using an Axio Imager 7.1 microscope (Zeiss) and images were taken using an Axio Cam MRm camera (Zeiss). The immunofluorescence stained slides were also examined by a confocal laser scanning microscope (CLSM) (Leica CTR 6500, Heidelberg). Images were processed using Photoshop.
ab32118 showing positive staining in Breast carcinoma tissue.
All lanes: Western blot - Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/10000 dilution
All lanes: A431 cell lysate
Predicted band size: 48 kDa
Observed band size: 48 kDa
ab32118 showing positive staining in Gastric adenocarcinoma tissue.
ab32118 showing positive staining in Normal colon tissue.
ab32118 showing positive staining in Normal kidney tissue.
ab32118 showing negative staining in Glioma tissue.
ab32118 showing negative staining in Normal brain tissue.
Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney tissue using ab32118. Green-CK18 red-PI
Overlay histogram showing MCF7 cells stained with ab32118 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32118, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab32118 at 1/20 dilution (2μg/ml) (red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluorr® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
R-PE (PE Anti-Cytokeratin 18 antibody [E431-1] ab210410) conjugated version is available for this clone.
Flow cytometry overlay histogram showing left MCF7 positive cells and right negative A375 stained with ab32118 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab32118) (1x 106 in 100μl at 0.008μg/ml (1/258750)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in MCF7 Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
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