Overview

  • Product name

    Anti-Cytokeratin 14 antibody [EP1612Y]
    See all Cytokeratin 14 primary antibodies
  • Description

    Rabbit monoclonal [EP1612Y] to Cytokeratin 14
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cytokeratin 14 aa 400-500 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: A431 cell lysate. IHC-P: Human skin tissue. ICC/IF: A431 cells. Flow Cyt: A431 cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP1612Y
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab51054 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
WB 1/20000. Detects a band of approximately 48 kDa (predicted molecular weight: 52 kDa).
IP 1/50.
Flow Cyt 1/100.

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

IHC-P Use at an assay dependent concentration.
ICC/IF 1/100.

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

  • ab51054 staining Cytokeratin 14 in human skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer, pH 6.0. Samples were then permeabilized using 0.1% saponin/PBS, blocked with 4% BSA for 30 minutes at 25°C and then incubated with ab51054 at a 1/200 dilution for 16 hours at 4°C. The secondary used was a Texas Red conjugated goat anti-rabbit polyclonal used at a 1/100 dilution.

    See Abreview

  • ICC/IF image of ab51504 stained A431 (Human epidermoid carcinoma cell line) cells.

    The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51504, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

  • Anti-Cytokeratin 14 antibody [EP1612Y] (ab51054) at 1/20000 dilution + A431 (Human epidermoid carcinoma cell line) cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit-HRP at 1/2000 dilution

    Predicted band size: 52 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?

  • Overlay histogram showing A431 (Human epidermoid carcinoma cell line) cells stained with ab51054 (red line).

    The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab51054, 1/100 dilution ) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was was rabbit IgG (monoclonal) ( 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.

References

This product has been referenced in:

  • Deng H  et al. Establishment and optimization of epithelial cell cultures from human ectocervix, transformation zone, and endocervix optimization of epithelial cell cultures. J Cell Physiol 234:7683-7694 (2019). Read more (PubMed: 30609028) »
  • Ragle LE  et al. Long-label-retaining mammary epithelial cells are created early in ductal development and distributed throughout the branching ducts. Mech Dev 159:103565 (2019). Read more (PubMed: 31336167) »
See all 11 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Primary keratinocytes)
Gel Running Conditions
Reduced Denaturing (any kD BioRad)
Loading amount
50 µg
Specification
Primary keratinocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Julien Coutier

Verified customer

Submitted Jul 02 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Adult mammary skin)
Permeabilization
No
Specification
Adult mammary skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C

Mr. Julien Coutier

Verified customer

Submitted Jun 14 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Pt link 3 in 1
Permeabilization
Yes - TBS-Tween 0.01%
Specification
skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 24°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 28 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Sample
Human Cell (Keratinocytes)
Specification
Keratinocytes
Permeabilization
Yes - 0,1%Saponin / PBS
Fixative
Methanol

Abcam user community

Verified customer

Submitted Jul 18 2013

Application
Western blot
Sample
Human Tissue lysate - whole (skin biopsie)
Loading amount
15 µg
Specification
skin biopsie
Gel Running Conditions
Reduced Non-Denaturing (Native) (5-16% gel)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 21 2013

Application
Flow Cytometry
Sample
Human Cell (keratinocytes)
Specification
keratinocytes
Preparation
Cell harvesting/tissue preparation method: cells were isolated from skin biopsie followed by cell culture
Sample buffer: PBS/10% human IgG
Fixation
Paraformaldehyde
Permeabilization
Yes - 0,1% Tritonx-100/PBS
Gating Strategy
no gate

Abcam user community

Verified customer

Submitted Jan 21 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin)
Specification
skin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6,0
Permeabilization
Yes - 0,1% Saponin /PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 03 2012

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