Recombinant Anti-Cytokeratin 18 antibody [SP69] (ab93741)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP69] to Cytokeratin 18
- Suitable for: mIHC, WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cytokeratin 18 antibody [SP69]
See all Cytokeratin 18 primary antibodies -
Description
Rabbit monoclonal [SP69] to Cytokeratin 18 -
Host species
Rabbit -
Tested applications
Suitable for: mIHC, WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Cytokeratin 18 aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P05783 -
Positive control
- IHC-P: Human prostate or breast carcinoma, Human stomach, and Human endometrium tissue; Flow Cyt (Intra): HeLa cells; ICC/IF: HeLa cells. mIHC: Human breast.
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General notes
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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Purity
Protein A/G purified -
Purification notes
Purified from TCS by protein A/G. -
Clonality
Monoclonal -
Clone number
SP69 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab93741 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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mIHC |
Use at an assay dependent concentration.
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WB |
1/200. Predicted molecular weight: 48 kDa.
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IHC-P |
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Incubate for 30 minutes at RT. Perform heat mediated antigen retrieval before commencing with IHC staining protocol by boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.
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ICC/IF |
1/1000.
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Flow Cyt (Intra) |
1/2000.
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Notes |
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mIHC
Use at an assay dependent concentration. |
WB
1/200. Predicted molecular weight: 48 kDa. |
IHC-P
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Incubate for 30 minutes at RT. Perform heat mediated antigen retrieval before commencing with IHC staining protocol by boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min. |
ICC/IF
1/1000. |
Flow Cyt (Intra)
1/2000. |
Target
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Function
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. -
Tissue specificity
Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma. -
Involvement in disease
Defects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600]. -
Sequence similarities
Belongs to the intermediate filament family. -
Post-translational
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 localization
Cytoplasm > perinuclear region. - Information by UniProt
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Database links
- Entrez Gene: 3875 Human
- Omim: 148070 Human
- SwissProt: P05783 Human
- Unigene: 406013 Human
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Alternative names
- Cell proliferation inducing gene 46 protein antibody
- Cell proliferation inducing protein 46 antibody
- Cell proliferation-inducing gene 46 protein antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast labelling Cytokeratin 18 with ab93741 at 1/200 dilution (1.02 µg/mL) (B), Collagen VI with ab271938 at 1/500 dilution (2.084 μg/ml) (C) and Cytokeratin 14 with ab236439 at 1/2000 dilution ( 0.519 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Panel A: merged staining of anti-Cytokeratin 14 (magenta; Opal™690), anti-Cytokeratin 18 (green; Opal™520) and anti-Collagen VI (red; Opal™570) on human breast.
Panel B: anti-Cytokeratin 18 stained on luminal epithelial cells.
Panel C: anti-Collagen VI stained on stroma.
Panel D: anti-Cytokeratin 14 stained on myoepithelial cells.The section was incubated in three rounds of staining: in the order of ab236439 for 30 mins, ab93741 for 10 mins, and ab271938 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium tissue sections labeling Cytokeratin 18 with ab93741 at 1/200 dilution (1.21 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach tissue sections labeling Cytokeratin 18 with ab93741 at 1/200 dilution (1.21 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab93741 at 1:100(2.4 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab93741 at 1:240 dilution (1.00 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.
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ICC/IF image of ab93741 stained HCT116 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab93741 at 1/500 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti-Rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Immunihistochemical staining of human stomach with ab93741.
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Flow cytometric analysis of rabbit anti-Cytokeratin 18 (SP69) antibody ab93741(1/100) in HeLa cells (green) compared to negative control of rabbit IgG (blue).
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Overlay histogram showing MCF7 cells stained with ab93741 (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 (ab93741, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. 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.
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ab93741 at 1/200 dilution staining Cytokeratin 18 in formalin-fixed, paraffin-embedded Human breast carcinoma tissue.
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Anti-Cytokeratin 18 antibody [SP69] (ab93741) at 1/25 dilution + A431 cell lysate
Predicted band size: 48 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab93741 has been referenced in 7 publications.
- Charbonneau M et al. Establishment of a ccRCC patient-derived chick chorioallantoic membrane model for drug testing. Front Med (Lausanne) 9:1003914 (2022). PubMed: 36275794
- Chin A et al. miR-1227 Targets SEC23A to Regulate the Shedding of Large Extracellular Vesicles. Cancers (Basel) 13:N/A (2021). PubMed: 34831007
- Mariscal J et al. Comprehensive palmitoyl-proteomic analysis identifies distinct protein signatures for large and small cancer-derived extracellular vesicles. J Extracell Vesicles 9:1764192 (2020). PubMed: 32944167
- Vagner T et al. Large extracellular vesicles carry most of the tumour DNA circulating in prostate cancer patient plasma. J Extracell Vesicles 7:1505403 (2018). PubMed: 30108686
- Schütz B et al. Chemical coding and chemosensory properties of cholinergic brush cells in the mouse gastrointestinal and biliary tract. Front Physiol 6:87 (2015). PubMed: 25852573
- Xiang J et al. CXCR4 Protein Epitope Mimetic Antagonist POL5551 Disrupts Metastasis and Enhances Chemotherapy Effect in Triple-Negative Breast Cancer. Mol Cancer Ther 14:2473-85 (2015). PubMed: 26269605
- Rodrigues RM et al. Human skin-derived stem cells as a novel cell source for in vitro hepatotoxicity screening of pharmaceuticals. Stem Cells Dev 23:44-55 (2014). PubMed: 23952781