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  1. Link

    cytokeratin-13-antibody-ae8-ab16112.pdf

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Tags & Cell Markers Cell Type Markers Epi / Endo-thelial
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Validated using a knockout cell line

Anti-Cytokeratin 13 antibody [AE8] (ab16112)

  • Datasheet
  • SDS
Reviews (3)Q&A (1)References (20)

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Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
  • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
  • Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)

Key features and details

  • Mouse monoclonal [AE8] to Cytokeratin 13
  • Suitable for: IHC-Fr, ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Cytokeratin 13 antibody [AE8]
    See all Cytokeratin 13 primary antibodies
  • Description

    Mouse monoclonal [AE8] to Cytokeratin 13
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-Fr, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rabbit
  • Immunogen

    Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431 whole cell lysate. ICC/IF: A431 cells. IHC-P: Human tonsil. IHC-Fr: Human tonsil.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Primary antibody notes

    This antibody is specific for Cytokeratin 13, which is a marker for oesophageal type differentiation which is expressed by various internal stratified epithelia.
  • Clonality

    Monoclonal
  • Clone number

    AE8
  • Myeloma

    P3-X63 Ag8.3
  • Isotype

    IgG
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Cell Type Markers
    • Epi / Endo-thelial
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class I
    • Cytokeratins

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG - Isotype Control (ab37355)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab16112 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 a concentration of 1 µg/ml.
ICC/IF (1)
Use a concentration of 1 - 5 µg/ml.

PubMed: 25076852

WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 50 kDa.
IHC-P (2)
Use a concentration of 0.05 µg/ml.
Notes
IHC-Fr
Use a concentration of 1 µg/ml.
ICC/IF
Use a concentration of 1 - 5 µg/ml.

PubMed: 25076852

WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 50 kDa.
IHC-P
Use a concentration of 0.05 µg/ml.

Target

  • Tissue specificity

    Expressed in some epidermal sweat gland ducts (at protein level) and in exocervix, esophagus and placenta.
  • Involvement in disease

    Defects in KRT13 are a cause of white sponge nevus of cannon (WSN) [MIM:193900]. WSN is a rare autosomal dominant disorder which predominantly affects non-cornified stratified squamous epithelia. Clinically, it is characterized by the presence of soft, white, and spongy plaques in the oral mucosa. The characteristic histopathologic features are epithelial thickening, parakeratosis, and vacuolization of the suprabasal layer of oral epithelial keratinocytes. Less frequently the mucous membranes of the nose, esophagus, genitalia and rectum are involved.
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Post-translational
    modifications

    O-glycosylated; glycans consist of single N-acetylglucosamine residues.
  • Target information above from: UniProt accession P13646 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3860 Human
    • Entrez Gene: 16663 Mouse
    • Omim: 148065 Human
    • SwissProt: P13646 Human
    • SwissProt: P08730 Mouse
    • Unigene: 654550 Human
    • Unigene: 4646 Mouse
    • Alternative names

      • 47 kDa cytokeratin antibody
      • CK-13 antibody
      • CK13 antibody
      • Cytokeratin 13 antibody
      • Cytokeratin-13 antibody
      • K13 antibody
      • K1C13_HUMAN antibody
      • Ka13 antibody
      • Keratin 13 antibody
      • Keratin antibody
      • keratin type I cytoskeletal 13 antibody
      • Keratin-13 antibody
      • Krt-1.13 antibody
      • Krt1-13 antibody
      • KRT13 antibody
      • MGC161462 antibody
      • MGC3781 antibody
      • type I cytoskeletal 13 antibody
      • Type I keratin Ka13 antibody
      • WSN2 antibody
      see all

    Images

    • Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      All lanes : Anti-Cytokeratin 13 antibody [AE8] (ab16112) at 1 µg/ml

      Lane 1 : Wild-type A431 cell lysate
      Lane 2 : KRT13 knockout A431 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 50 kDa
      Observed band size: 51 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-Cytokeratin 13 antibody [AE8] staining at 1 µg/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab16112 was shown to bind specifically to Cytokeratin 13. A band was observed at 51 kDa in wild-type A431 cell lysates with no signal observed at this size in Krt13 knockout cell line ab269483 (knockout cell lysate ab269647). To generate this image, wild-type and Krt13 knockout A431 cell lysates were analysed. 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)

      IHC image of Cytokeratin 13 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F.

      The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16112, 0.05 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)

      IHC image of Cytokeratin 13 staining in a section of frozen normal human tonsil*. The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab16112 at 1µg/ml. The section was then incubated with ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488), 1/1000)) (shown in green) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. The secondary-only control insert image is taken from an identical assay without primary antibody. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

    • Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Anti-Cytokeratin 13 antibody [AE8] (ab16112) at 1 µg/ml + A431 whole cell lysate at 20 µg

      Performed under reducing conditions.

      Predicted band size: 50 kDa



      This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab16112 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 13 antibody [AE8] (ab16112)

      ab16112 staining Cytokeratin 13 in A431 cells. The cells were fixed with Methanol (5min), permeabilized with 0.1%PBS-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 ab16112 at 5μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

       

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)This image is courtesy of an anonymous Abreview

      ab16112 staining Cytokeratin 13 in Human pharynx tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in Tris pH9. Samples were incubated with primary antibody (undiluted) for 1 hour at 20°C. An undiluted HRP-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody.

      See Abreview

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (20)

    Publishing research using ab16112? Please let us know so that we can cite the reference in this datasheet.

    ab16112 has been referenced in 20 publications.

    • Uetsuki R  et al. The squamous cell carcinoma cell line OM-1 retains both p75-dependent stratified epithelial progenitor potential and cancer stem cell properties. Biochem Biophys Rep 26:101003 (2021). PubMed: 34041369
    • Konishi T  et al. LRRC8A Expression Influences Growth of Esophageal Squamous Cell Carcinoma. Am J Pathol 189:1973-1985 (2019). PubMed: 31323188
    • Schmitt L  et al. Novel Human Full-Thickness Three-Dimensional Nonkeratinized Mucous Membrane Model for Pharmacological Studies in Wound Healing. Skin Pharmacol Physiol 32:265-274 (2019). PubMed: 31284289
    • Nakano K  et al. Notch Signaling Affects Oral Neoplasm Cell Differentiation and Acquisition of Tumor-Specific Characteristics. Int J Mol Sci 20:N/A (2019). PubMed: 31018488
    • Sakima VT  et al. Antimicrobial Photodynamic Therapy Mediated by Curcumin-Loaded Polymeric Nanoparticles in a Murine Model of Oral Candidiasis. Molecules 23:N/A (2018). PubMed: 30126245
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Immunocytochemistry/ Immunofluorescence abreview for Anti-Cytokeratin 13 antibody [AE8]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Lip)
    Specification
    Lip
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

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    Submitted Aug 14 2020

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