Recombinant Anti-Cytokeratin 20 antibody [EPR1622Y] - BSA and Azide free (ab219589)
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 - BSA and Azide free
- 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] - BSA and Azide free
See all Cytokeratin 20 primary antibodies -
Description
Rabbit monoclonal [EPR1622Y] to Cytokeratin 20 - BSA and Azide free -
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.
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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 whole 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
ab219589 is the carrier-free version of ab76126.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 3.10 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
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)
- Alexa Fluor® 488 Anti-Cytokeratin 20 antibody [EPR1622Y] (ab275988)
- Alexa Fluor® 555 Anti-Cytokeratin 20 antibody [EPR1622Y] (ab280839)
- Alexa Fluor® 594 Anti-Cytokeratin 20 antibody [EPR1622Y] (ab303597)
- Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (ab76126)
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab219589 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) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa. |
IHC-P
Use at an assay dependent concentration. 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
Observed band size: 45-50 kDa why is the actual band size different from the predicted?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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
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ab76126 at 1/100 dilution staining Cytokeratin 20 in human colon adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
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ab76126 showing positive staining in human colonic adenocarcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab76126 showing negative staining in human hepatocellular carcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab76126 showing negative staining in human breast carcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76126).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (7)
ab219589 has been referenced in 7 publications.
- Hayashi M et al. The adenosine A2B receptor is involved in anion secretion in human pancreatic duct Capan-1 epithelial cells. Pflugers Arch 468:1171-81 (2016). IHC-P ; Rat . PubMed: 26965147
- Lee J et al. Walnut Phenolic Extract and Its Bioactive Compounds Suppress Colon Cancer Cell Growth by Regulating Colon Cancer Stemness. Nutrients 8:N/A (2016). WB ; Human . PubMed: 27455311
- Strand DW et al. Deficiency in metabolic regulators PPAR? and PTEN cooperates to drive keratinizing squamous metaplasia in novel models of human tissue regeneration. Am J Pathol 182:449-59 (2013). WB . PubMed: 23219716
- Zeestraten EC et al. Specific activity of cyclin-dependent kinase I is a new potential predictor of tumour recurrence in stage II colon cancer. Br J Cancer 106:133-40 (2012). ICC/IF ; Human . PubMed: 22108518
- Volkmer JP et al. Three differentiation states risk-stratify bladder cancer into distinct subtypes. Proc Natl Acad Sci U S A 109:2078-83 (2012). IHC-P ; Human . PubMed: 22308455
- Vrang N et al. The effects of 13 wk of liraglutide treatment on endocrine and exocrine pancreas in male and female ZDF rats: a quantitative and qualitative analysis revealing no evidence of drug-induced pancreatitis. Am J Physiol Endocrinol Metab 303:E253-64 (2012). IHC-P ; Rat . PubMed: 22589391
- Gulhati P et al. mTORC1 and mTORC2 Regulate EMT, Motility, and Metastasis of Colorectal Cancer via RhoA and Rac1 Signaling Pathways. Cancer Res 71:3246-56 (2011). IHC-P ; Human . PubMed: 21430067