Recombinant
RabMAb

Anti-Cytokeratin 20 antibody [EPR1622Y] (ab76126)

Overview

  • Product name
    Anti-Cytokeratin 20 antibody [EPR1622Y]
    See all Cytokeratin 20 primary antibodies
  • Description
    Rabbit monoclonal [EPR1622Y] to Cytokeratin 20
  • 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: IHC-Fr, ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Rat, Goat, Human, Pig, Common marmoset
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cytokeratin 20 (C terminal).

  • Positive control
    • Human small intestine lysate; human colon adenocarcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Alternative versions available:

    Anti-Cytokeratin 20 antibody (HRP) [EPR1622Y] (ab194220)

    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

Applications

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
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/100 - 1/500.
WB 1/10000 - 1/50000. Predicted molecular weight: 48 kDa.
IP 1/40.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • 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
    modifications
    Hyperphosphorylation 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
  • Database links
  • Alternative names
    • CD20 antibody
    • CK 20 antibody
    • CK-20 antibody
    • CK20 antibody
    • Cytokeratin-20 antibody
    • Cytokeratin20 antibody
    • K1C20_HUMAN antibody
    • K20 antibody
    • KA20 antibody
    • Keratin 20 antibody
    • keratin 20, type I antibody
    • keratin 21, rat, homolog of antibody
    • Keratin antibody
    • Keratin type I cytoskeletal 20 antibody
    • Keratin-20 antibody
    • Keratin20 antibody
    • KRT 20 antibody
    • KRT 21 antibody
    • KRT20 antibody
    • KRT21 antibody
    • MGC35423 antibody
    • OTTHUMP00000164518 antibody
    • Protein IT antibody
    • type I cytoskeletal 20 antibody
    see all

Images

  • 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.

  • Anti-Cytokeratin 20 antibody [EPR1622Y] (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

  • ab76126 at 1/100 dilution staining Cytokeratin 20 in human colon adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.
  • Overlay histogram showing LoVo 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.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 48 kDa


    Exposure time : 20 seconds

    Primary: ab76126, 1/50000 dilution
    Sample: ab73640, 0.01 ug
    Secondary: ab97080, 1/5000 dilution

  • ab76126 showing positive staining in Colonic adenocarcinoma tissue.

  • ab76126 showing positive staining in Urinary bladder transitional carcinoma tissue.

  • ab76126 showing negative staining in Hepatocellular carcinoma tissue.

  • ab76126 showing negative staining in Breast carcinoma tissue.

  • Fluorescent immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using ab76126. Green-CK20 red-PI

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Lou Y  et al. Estradiol Suppresses TLR4-triggered Apoptosis of Decidual Stromal Cells and Drives an Anti-inflammatory TH2 Shift by Activating SGK1. Int J Biol Sci 13:434-448 (2017). Read more (PubMed: 28529452) »
  • Kuracha MR  et al. Patient-derived xenograft mouse models of pseudomyxoma peritonei recapitulate the human inflammatory tumor microenvironment. Cancer Med 5:711-9 (2016). IHC . Read more (PubMed: 26833741) »

See all 12 Publications for this product

Customer reviews and Q&As

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You may have to permeabilize the sections with a buffer stronger than tween to improve staining, such as triton x-100. This may allow the antibody greater access to the cytoplasmic p...

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I am sorry to hear you have had difficulty obtaining satisfactory results from this Cytokeratin 20 antibody.

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