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Anti-Cytokeratin 18 [C-04] antibody (ab668)抗細胞角蛋白18抗體產品信息:
Mouse monoclonal [C-04] to Cytokeratin 18
Reacts with: Mouse, Rat, Sheep, Goat, Horse, Hamster, Cow, Cat, Dog, Human, Pig, Marmoset (common)
Tissue / cell preparation (Human). (Cytoskeleton preparation of epidermal carcinoma cell line A431).
Properties
Concentration 100 µg at 1 mg/ml
Applications
Our Abpromise guarantee covers the use of ab668 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Notes |
---|---|
ICC | ICC: Use at an assay dependent dilution. |
IHC-P | IHC-P: Use at an assay dependent dilution. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB | WB: Use at an assay dependent dilution. Predicted molecular weight: 45 kDa. |
IHC-FoFr | IHC-FoFr: Use at an assay dependent concentration. |
Flow Cyt | Flow Cyt: Use 1µg for 106 cells. (also see PMID 18946470) |
IHC-Fr | IHC-Fr: Use at an assay dependent dilution. |
IP | IP: Use at an assay dependent dilution. |
ICC/IF | ICC/IF: Use at an assay dependent dilution. |
Target
Database links
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Alternative names
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Anti-Cytokeratin 18 [C-04] antibody images
Flow Cytometry - Anti-Cytokeratin 18 antibody [C-04] (ab668)
Overlay histogram showing HCT116 cells stained with ab668 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab668, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HCT116 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.
Immunocytochemistry/ Immunofluorescence - Cytokeratin 18 antibody [C-04] (ab668)This image is courtesy of an anonymous Abreview
ab668 at 1/100 staining A431 Human epithelial carcinoma cells by ICC/IF. The cells were paraformaldehyde fixed, permeabilized with 0.25% Triton X-100 and then blocked with BSA prior to incubation with the antibody for 24 hours at 4°C. A FITC conjugated goat anti-mouse IgG antibody was used as the secondary. The right hand image shows staining with ab668 followed by secondary antibody. The left hand image shows DAPI staining with secondary only.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytokeratin 18 antibody [C-04] (ab668)
ab668 (2µg/ml) staining cytokeratin 18 in human skin using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of sweat coils.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 [C-04] antibody (ab668)This image is courtesy of an Abreview submitted by Carl Hobbs
ab668 staining Cytokeratin 18 in Cat lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/500 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.
Coloured arrowheads indicate positivity.
Good immunolabelling of bronchial tree epithelia (green), alveolar lining epithelium (red ).
Goblet cells are negative (asterisk).
Heavily stained structures are submucosal mucous glands.
Immunocytochemistry/ Immunofluorescence - Cytokeratin 18 antibody [C-04] (ab668)Image courtesy of an anonymous Abreview.
ab668 staining Cytokeratin 18 in horse primary bronchial epithelial cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in acetone and then blocked using 3% BSA for 10 hours at 4°C. Samples were then incubated with primary antibody at 1/100 for 1 hour at 22°C. The secondary antibody used was a goat anti-mouse conjugated to FITC used at a 1/200 dilution.
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