Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)
![Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-11-anti-cytokeratin-8-antibody-m20-western-blot-human-lysate.jpg "Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)")
Key features and details
- Mouse monoclonal [M20] to Cytokeratin 8 - Cytoskeleton Marker
- Suitable for: Flow Cyt, IHC-P, IHC-Fr, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker
See all Cytokeratin 8 primary antibodies -
Description
Mouse monoclonal [M20] to Cytokeratin 8 - Cytoskeleton Marker -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanIHC-Fr HumanIHC-P HumanWB Human
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Immunogen
Keratin isolated from the human breast carcinoma cell line MCF-7.
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Positive control
- WB: HeLa, A431, PC3 and LNCaP cell lysates. ICC/IF: MCF-7, pancreatic cancer and epithelial ovarian cancer cells. IHC-Fr: Human colon tissue.
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General notes
Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 – 7.8. The individual cytokeratin polypeptides are numbered 1 to 20. The various epithelia in the human body usually express cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. The cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.3
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% Fetal calf serum -
Concentration information loading... -
Purity
Tissue culture supernatant -
Primary antibody notes
Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 – 7.8. The individual cytokeratin polypeptides are numbered 1 to 20. The various epithelia in the human body usually express cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. The cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays. -
Clonality
Monoclonal -
Clone number
M20 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab9023 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| Flow Cyt |
Human
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| IHC-Fr |
Human
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| IHC-P |
Human
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| WB |
Human
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| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt |
Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| IHC-P | (1) |
Use a concentration of 4 µg/ml.
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| IHC-Fr |
1/100 - 1/200. For human colon 20min ice cold acetone fixation was carried out with 60min incubation with ab9023 at 37C.
|
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| WB | (4) |
1/100 - 1/1000.
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| Notes |
|---|
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Flow Cyt
Use 2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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IHC-P
Use a concentration of 4 µg/ml. |
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IHC-Fr
1/100 - 1/200. For human colon 20min ice cold acetone fixation was carried out with 60min incubation with ab9023 at 37C. |
|
WB
1/100 - 1/1000. |
Target
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Function
Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle. -
Tissue specificity
Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma membrane in structures that contain dystrophin and spectrin. Expressed in gingival mucosa and hard palate of the oral cavity. -
Involvement in disease
Cirrhosis -
Sequence similarities
Belongs to the intermediate filament family. -
Post-translational
modificationsPhosphorylation on serine residues is enhanced during EGF stimulation and mitosis. Ser-74 phosphorylation plays an important role in keratin filament reorganization.
O-glycosylated. O-GlcNAcylation at multiple sites increases solubility, and decreases stability by inducing proteasomal degradation.
O-glycosylated (O-GlcNAcylated), in a cell cycle-dependent manner. -
Cellular localization
Cytoplasm. Nucleus, nucleoplasm. Nucleus matrix. - Information by UniProt
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Database links
- Entrez Gene: 3856 Human
- Omim: 148060 Human
- SwissProt: P05787 Human
- Unigene: 533782 Human
- Unigene: 708445 Human
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Alternative names
- CARD2 antibody
- CK 8 antibody
- CK-8 antibody
see all
Images
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Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)All lanes : Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)
Lane 1 : A431 cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : KRT8 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.Lanes 1 - 4: Merged signal (red and green). Green - ab9023 observed at 55 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab9023 was shown to react with Cytokeratin 8 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255400 (knockout cell lysate ab263785) was used. Wild-type and Cytokeratin 8 knockout samples were subjected to SDS-PAGE. ab9023 and Anti-GAPDH antibody EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-16015-anti-cytokeratin-8-antibody-m20-western-blot.jpg)
Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)The image shows different concentration of total protein from normal and tumour colon tissues.
This picture was kindly supplied as part of the review submitted by Semona Rupchand.
The image shows different concentration of total protein from normal and tumour colon tissues.
This picture was kindly supplied as part of the review submitted by Semona Rupchand.
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![Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-16011-anti-cytokeratin-8-antibody-m20-western-blot.jpg)
Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)This image is a courtesy of Anonymous AbreviewAll lanes : Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023) at 1/2000 dilution
Lane 1 : Lysate prepared from human prostate cancer LNCaP cells
Lane 2 : Lysate prepared from human prostate cancer PC3 cells
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : HRP-conjugated goat polyclonal to mouse IgG at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 50 kDa why is the actual band size different from the predicted?
Additional bands at: 26 kDa, 65 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
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![Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-457939-anti-cytokeratin-8-antibody-m20-cytoskeleton-marker-immunohistochemistry-frozen-colon-human.jpg)
Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)Immunohistochemistry analysis of frozen section of human colon labeling Cytokeratin 8 with ab9023.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-16008-anti-cytokeratin-8-antibody-m20-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)Immunohistochemistry on paraffin section of human colon -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-16012-anti-cytokeratin-8-antibody-m20-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)Ab9023 staining human normal liver parenchima tissue. Staining is localized to cell membrane.
Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required. -
![Flow Cytometry - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-16010-anti-cytokeratin-8-antibody-m20-flow-cytometry.jpg)
Flow Cytometry - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)Overlay histogram showing MCF-7 cells stained with ab9023 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.5% 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 (ab9023, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/250 dilution for 30 min at 22ºC. 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 MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.5% PBS-Tween used under the same conditions.
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![Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)](https://www.abcam.com/ps/products/9/ab9023/Images/ab9023-16016-anti-cytokeratin-8-antibody-m20-immunohistochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)Immunohistochemistry on frozen section of human colon
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (24)
ab9023 has been referenced in 24 publications.
- Cao J et al. Estrogen attenuates TGF-ß1-induced EMT in intrauterine adhesion by activating Wnt/ß-catenin signaling pathway. Braz J Med Biol Res 53:e9794 (2020). PubMed: 32638833
- Ehsani E et al. Improved differentiation of human enriched CD133+CD24+ renal progenitor cells derived from embryonic stem cell with embryonic mouse kidney-derived mesenchymal stem cells co-culture. Differentiation 109:1-8 (2019). PubMed: 31323479
- Nguyen QH et al. Profiling human breast epithelial cells using single cell RNA sequencing identifies cell diversity. Nat Commun 9:2028 (2018). PubMed: 29795293
- Baek A et al. Autophagy and KRT8/keratin 8 protect degeneration of retinal pigment epithelium under oxidative stress. Autophagy 13:248-263 (2017). PubMed: 28045574
- McCormick NH et al. ZnT4 (SLC30A4)-null ("lethal milk") mice have defects in mammary gland secretion and hallmarks of precocious involution during lactation. Am J Physiol Regul Integr Comp Physiol 310:R33-40 (2016). WB ; Mouse . PubMed: 26538236