Product nameAnti-Cytokeratin 18 antibody
See all Cytokeratin 18 primary antibodies
DescriptionRabbit polyclonal to Cytokeratin 18
Tested applicationsSuitable for: ICC/IF, Flow Cyt, IP, IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
- Hela, Jurkat and A431 cell lysates
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab24561 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).|
FunctionInvolved 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.
Tissue specificityExpressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.
Involvement in diseaseDefects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600].
Sequence similaritiesBelongs to the intermediate filament family.
modificationsPhosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6.
Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.
O-glycosylated at multiple sites; glycans consist of single N-acetylglucosamine residues.
Cellular localizationCytoplasm > perinuclear region.
- Information by UniProt
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ICC/IF image of ab24561 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab24561, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
All lanes : Anti-Cytokeratin 18 antibody (ab24561) at 1 µg/ml
Lane 1 : HeLa whole cell lysate
Lane 2 :
Jurkat whole cell lysate (ab7899)
Lane 3 :
A431 whole cell lysate (ab7909)
Lysates/proteins at 20 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 48 kDa
Observed band size: 48 kDa
ab24561 (1/2000) staining Cytokeratin 18 in paraffin-embedded Human liver tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.
Overlay histogram showing MCF7 cells stained with ab24561 (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 (ab24561, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (polyclonal) (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.
Cytokeratin 18 was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to Cytokeratin 18 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, A431 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab24561.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 48kDa; Cytokeratin 18