Overview

  • Product name

    Anti-Cytokeratin 14 antibody [EPR17350] - Cytoskeleton Marker
    See all Cytokeratin 14 primary antibodies
  • Description

    Rabbit monoclonal [EPR17350] to Cytokeratin 14 - Cytoskeleton Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse Cytokeratin 14 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q61781

  • Positive control

    • WB: A431 and HaCaT whole cell lysates; Mouse, rat and human skin lysates. IHC-P: Human skin and squamous cell carcinoma of cervix tissues; Mouse and rat skin tissues. ICC/IF: PC-12 cells. FC: PC-12 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17350
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab181595 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-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/20000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
ICC/IF 1/1000.
Flow Cyt 1/190.

Target

  • Function

    The nonhelical tail domain is involved in promoting KRT5-KRT14 filaments to self-organize into large bundles and enhances the mechanical properties involved in resilience of keratin intermediate filaments in vitro.
  • Tissue specificity

    Detected in the basal layer, lowered within the more apically located layers specifically in the stratum spinosum, stratum granulosum but is not detected in stratum corneum. Strongly expressed in the outer root sheath of anagen follicles but not in the germinative matrix, inner root sheath or hair. Found in keratinocytes surrounding the club hair during telogen.
  • Involvement in disease

    Defects in KRT14 are a cause of epidermolysis bullosa simplex Dowling-Meara type (DM-EBS) [MIM:131760]. DM-EBS is a severe form of intraepidermal epidermolysis bullosa characterized by generalized herpetiform blistering, milia formation, dystrophic nails, and mucous membrane involvement.
    Defects in KRT14 are a cause of epidermolysis bullosa simplex Weber-Cockayne type (WC-EBS) [MIM:131800]. WC-EBS is a form of intraepidermal epidermolysis bullosa characterized by blistering limited to palmar and plantar areas of the skin.
    Defects in KRT14 are a cause of epidermolysis bullosa simplex Koebner type (K-EBS) [MIM:131900]. K-EBS is a form of intraepidermal epidermolysis bullosa characterized by generalized skin blistering. The phenotype is not fundamentally distinct from the Dowling-Meara type, although it is less severe.
    Defects in KRT14 are the cause of epidermolysis bullosa simplex autosomal recessive (AREBS) [MIM:601001]. AREBS is an intraepidermal epidermolysis bullosa characterized by localized blistering on the dorsal, lateral and plantar surfaces of the feet.
    Defects in KRT14 are the cause of Naegeli-Franceschetti-Jadassohn syndrome (NFJS) [MIM:161000]; also known as Naegeli syndrome. NFJS is a rare autosomal dominant form of ectodermal dysplasia. The cardinal features are absence of dermatoglyphics (fingerprints), reticular cutaneous hyperpigmentation (starting at about the age of 2 years without a preceding inflammatory stage), palmoplantar keratoderma, hypohidrosis with diminished sweat gland function and discomfort provoked by heat, nail dystrophy, and tooth enamel defects.
    Defects in KRT14 are the cause of dermatopathia pigmentosa reticularis (DPR) [MIM:125595]. DPR is a rare ectodermal dysplasia characterized by lifelong persistent reticulate hyperpigmentation, noncicatricial alopecia, and nail dystrophy.
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Cellular localization

    Cytoplasm. Nucleus. Expressed in both as a filamentous pattern.
  • Information by UniProt
  • Database links

  • Alternative names

    • CK 14 antibody
    • CK-14 antibody
    • ck14 antibody
    • Cytokeratin 14 antibody
    • Cytokeratin-14 antibody
    • Cytokeratin14 antibody
    • Dowling Meara antibody
    • EBS3 antibody
    • EBS4 antibody
    • Epidermolysis bullosa simplex antibody
    • K14 antibody
    • K1C14_HUMAN antibody
    • Keratin 14 (epidermolysis bullosa simplex, Dowling-Meara, Koebner) antibody
    • Keratin 14 antibody
    • Keratin antibody
    • Keratin type I cytoskeletal 14 antibody
    • Keratin, type I cytoskeletal 14 antibody
    • Keratin-14 antibody
    • Keratin14 antibody
    • Koebner antibody
    • Krt 14 antibody
    • Krt14 antibody
    • NFJ antibody
    • OTTHUMP00000164624 antibody
    • type I cytoskeletal 14 antibody
    see all

Images

  • Immunohistochemistry of mouse SG (submandibular glands) epithelial cells.

    Immunohistochemical analysis of keratin 14 expression in normal SG tissue (SG) and P1 and P80 cell cultures.

    Scale bars = 10 μm.

    The  submandibular and sublingual glands are located together in the anterior neck between the submandibular lymph nodes and sternum. The white fat tissue and sublingual glands were removed to expose the SGs and main duct, which was ligated with a 4–0 nylon suture thread to ensure that only SG cells were isolated. The excised glands were embedded in optimal cutting temperature compound and frozen in liquid nitrogen. Samples stained with ab181595 at a 1:250 dilution 1% BSA in PBS overnight at 4°C.

  • Immunohistochemical analysis of paraffin-embedded human skin tissue labeling Cytokeratin 14 with ab181595 at 1/2000 dilution, followed by prediluted HRP polymer for Rabbit/Mouse IgG. Basal cells of epidermis show strong staining while no staining on the stratum corneum. Counterstained with hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted HRP polymer for Rabbit/Mouse IgG.

  • Immunofluorescent analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized PC-12 (Rat adrenal gland pheochromocytoma cells) labeling Cytokeratin 14 with ab181595 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on PC-12 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls:-
    -ve control 1: ab181595 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • All lanes : Anti-Cytokeratin 14 antibody [EPR17350] - Cytoskeleton Marker (ab181595) at 1/20000 dilution

    Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysate
    Lane 2 : HaCaT (Human keratinocyte cells) whole cell lysate
    Lane 3 : Mouse skin lysate
    Lane 4 : Rat skin lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    HaCaT cells express a higher level of Cytokeratin 14 than other cell lines such as A431.

  • Flow Cytometry analysis of PC-12 (rat adrenal gland pheochromocytoma) cells labeling Cytokeratin 14 with purified ab181595 at 1/190 dilution (10 µg/mL) (red).

    Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of cervix tissue labeling Cytokeratin 14 with ab181595 at 1/2000 dilution, followed by prediluted HRP polymer for Rabbit/Mouse IgG. Squamous carcinoma cells show strong staining. Counterstained with hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted HRP polymer for Rabbit/Mouse IgG.

  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling Cytokeratin 14 with ab181595 at 1/2000 dilution, followed by prediluted HRP polymer for Rabbit/Mouse IgG. Basal cells of epidermis show strong staining while no staining on the stratum corneum. Counterstained with hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted HRP polymer for Rabbit/Mouse IgG.

  • Immunohistochemical analysis of paraffin-embedded rat skin tissue labeling Cytokeratin 14 with ab181595 at 1/2000 dilution, followed by prediluted HRP polymer for Rabbit/Mouse IgG. Basal cells of epidermis show strong staining while no staining on the stratum corneum. Counterstained with hematoxylin.

    Negative control: PBS instead of primary antibody, secondary antibody is prediluted HRP polymer for Rabbit/Mouse IgG.

  • Anti-Cytokeratin 14 antibody [EPR17350] - Cytoskeleton Marker (ab181595) at 1/20000 dilution + Human skin lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Ghosh S  et al. Tumor Tissue Explant Culture of Patient-Derived Xenograft as Potential Prioritization Tool for Targeted Therapy. Front Oncol 9:17 (2019). Read more (PubMed: 30723707) »
  • Hu T  et al. Developing a Novel and Convenient Model for Investigating Sweat Gland Morphogenesis from Epidermal Stem Cells. Stem Cells Int 2019:4254759 (2019). Read more (PubMed: 30863451) »
See all 13 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (primary skin keratinicytes)
Gel Running Conditions
Reduced Denaturing (gel anyKd from BioRad)
Loading amount
15 µg
Specification
primary skin keratinicytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Mr. Julien Coutier

Verified customer

Submitted May 12 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Bladder cancer)
Permeabilization
Yes - 0.1% Triton X-100
Specification
Bladder cancer
Blocking step
Casein (DAKO ­ Protein Block Serum-Free) as blocking agent for 10 minute(s) · Concentration: 0.25% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 04 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
Sample
Mouse Tissue sections (mouse breast)
Specification
mouse breast
Permeabilization
Yes - Tween-20
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 04 2015

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