Recombinant Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (ab76126)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1622Y] to Cytokeratin 20 - Cytoskeleton Marker
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker
See all Cytokeratin 20 primary antibodies -
Description
Rabbit monoclonal [EPR1622Y] to Cytokeratin 20 - Cytoskeleton Marker -
Host species
Rabbit -
Specificity
The immunogen of this antibody is 73% homolog with Mouse-Cytokeratin 20. This antibody gives positive results for mouse samples in Western Blot only. Therefore we do not recommend this antibody for mouse samples and do not cover mouse with our Abpromise guarantee. -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Rat, Goat, Pig, Common marmoset -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HT-29, DLD-1 and Human small intestine cell lysates. IHC-P: Human colon adenocarcinoma and urinary bladder transitional carcinoma tissue. ICC: HT-29 cells. Flow Cyt (intra): LoVo cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Dissociation constant (KD)
KD = 3.10 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1622Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- PE Anti-Cytokeratin 20 antibody [EPR1622Y] (ab209923)
- Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (ab219589)
- Alexa Fluor® 488 Anti-Cytokeratin 20 antibody [EPR1622Y] (ab275988)
- Alexa Fluor® 647 Anti-Cytokeratin 20 antibody [EPR1622Y] (ab279694)
- Alexa Fluor® 555 Anti-Cytokeratin 20 antibody [EPR1622Y] (ab280839)
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76126 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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Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (3) |
1/100 - 1/500.
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WB |
1/10000 - 1/50000. Predicted molecular weight: 48 kDa.
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IHC-P | (12) |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/500. |
WB
1/10000 - 1/50000. Predicted molecular weight: 48 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
Plays a significant role in maintaining keratin filament organization in intestinal epithelia. When phosphorylated, plays a role in the secretion of mucin in the small intestine. -
Tissue specificity
Expressed predominantly in the intestinal epithelium. Expressed in luminal cells of colonic mucosa. Also expressed in the Merkel cells of keratinized oral mucosa; specifically at the tips of some rete ridges of the gingival mucosa, in the basal layer of the palatal mucosa and in the taste buds of lingual mucosa. -
Sequence similarities
Belongs to the intermediate filament family. -
Developmental stage
First detected at embryonic week 8 in individual 'converted' simple epithelial cells of the developing intestinal mucosa. In later fetal stages, synthesis extends over most goblet cells and a variable number of villus enterocytes. In the developing gastric and intestinal mucosa, expressed in all enterocytes and goblet cells as well as certain 'low-differentiated' columnar cells, whereas the neuroendocrine and Paneth cells are negative. -
Post-translational
modificationsHyperphosphorylation at Ser-13 occurs during the early stages of apoptosis but becomes less prominent during the later stages. Phosphorylation at Ser-13 also increases in response to stress brought on by cell injury.
Proteolytically cleaved by caspases during apoptosis. Cleavage occurs at Asp-228. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 54474 Human
- Entrez Gene: 286912 Rat
- Omim: 608218 Human
- SwissProt: P35900 Human
- SwissProt: P25030 Rat
- Unigene: 84905 Human
- Unigene: 9887 Rat
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Alternative names
- CD20 antibody
- CK 20 antibody
- CK-20 antibody
see all
Images
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All lanes : Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (ab76126) at 1/10000 dilution
Lane 1 : HT-29 cell lysate
Lane 2 : DLD-1 cell lysate
Lane 3 : SW480 cell lysate
Lane 4 : Daudi cell lysate
Lane 5 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 48 kDa
Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.False colour image of Western blot: Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76126 was shown to bind specifically to Cytokeratin 20. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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ab76126 staining Cytokeratin 20 in HT-29 cells, with negative expression in MCF7 cells. The cells were fixed with 100% methanol (5 min), permeabilised 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. The cells were then incubated overnight at +4°C with ab76126 at 0.1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 µg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
This product also work with 4% formaldehyde (10 min) fixation under the same testing conditions.
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ab76126 at 1/100 dilution staining Cytokeratin 20 in human colon adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) cells labelling Cytokeratin 20 with purified ab76126 at 1/500. Cells were fixed with 100% methanol. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
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Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (ab76126) at 1/50000 dilution + human small intestine lysate at 10 µg
Secondary
goat anti-rabbit-HRP at 1/1000 dilution
Predicted band size: 48 kDa
Observed band size: 44 kDa why is the actual band size different from the predicted?Primary: ab76126, 1/50000 dilution
Sample: human small intestine lysate, 10 ug
Secondary: goat anti-rabbit-HRP, 1/1000 dilution -
Overlay histogram showing LoVo (Human colorectal adenocarcinoma cell line) cells stained with ab76126 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab76126, 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) (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.
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Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (ab76126) at 1/50000 dilution + Recombinant Human Cytokeratin 20 protein (ab73640) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 48 kDa
Exposure time: 20 secondsPrimary: ab76126, 1/50000 dilution
Sample: ab73640, 0.01 ug
Secondary: ab97080, 1/5000 dilution -
ab76126 showing positive staining in human colonic adenocarcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab76126 showing positive staining in human urinary bladder transitional carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab76126 showing negative staining in human hepatocellular carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab76126 showing negative staining in human breast carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Fluorescent immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using ab76126. Green-CK20 red-PI
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (46)
ab76126 has been referenced in 46 publications.
- Jhang JF et al. Improved Urothelial Cell Proliferation, Cytoskeleton and Barrier Function Protein Expression in the Patients With Interstitial Cystitis/Bladder Pain Syndrome After Intravesical Platelet-Rich Plasma Injection. Int Neurourol J 26:S57-67 (2022). PubMed: 35073671
- Wu SY et al. Inflammation and Barrier Function Deficits in the Bladder Urothelium of Patients with Chronic Spinal Cord Injury and Recurrent Urinary Tract Infections. Biomedicines 10:N/A (2022). PubMed: 35203430
- Shibata O et al. Establishment of a pancreatic cancer animal model using the pancreas-targeted hydrodynamic gene delivery method. Mol Ther Nucleic Acids 28:342-352 (2022). PubMed: 35474735
- Wang X et al. Disulfiram Exerts Antifibrotic and Anti-Inflammatory Therapeutic Effects on Perimysial Orbital Fibroblasts in Graves' Orbitopathy. Int J Mol Sci 23:N/A (2022). PubMed: 35563653
- Forsythe SD et al. Application of immune enhanced organoids in modeling personalized Merkel cell carcinoma research. Sci Rep 12:13865 (2022). PubMed: 35974123