Anti-Cytokeratin 14 antibody [LL002] (ab7800)

Mouse monoclonal Cytokeratin 14 antibody [LL002]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 88 publication(s). Independently reviewed in 18 review(s).

Overview

  • Product name
    Anti-Cytokeratin 14 antibody [LL002]
    See all Cytokeratin 14 primary antibodies
  • Description
    Mouse monoclonal [LL002] to Cytokeratin 14
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IF, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Cytokeratin 14 (C terminal).
    Database link: P02533

  • Positive control
    • IHC-P: Human normal skin tissue sections. ICC/IF: A431 cells. WB: A431 whole cell lysate. Human skin whole tissue lysate
  • General notes

    This antibody labels the basal layer of stratifying squamous and non-squamous epithelia. The staining pattern iscytoplasmic. It recognizes basal cell carcinomas and squamous cell carcinomas.

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

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot. Store at +4°C. Do Not Freeze.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity
    Protein A purified
  • Primary antibody notes
    This antibody labels the basal layer of stratifying squamous and non-squamous epithelia. The staining pattern iscytoplasmic. It recognizes basal cell carcinomas and squamous cell carcinomas.
  • Clonality
    Monoclonal
  • Clone number
    LL002
  • Isotype
    IgG3
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7800 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 Use a concentration of 1 µg/ml.
Flow Cyt Use at an assay dependent concentration.

0.5 - 1 µg/million cells in 0.1 ml.

ab91537 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 0.1 - 1 µg/ml.
IHC-P Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr 1/20 - 1/300. PubMed: 23769181

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

  • IHC image of Cytokeratin 14 staining in a section of formalin-fixed paraffin-embedded normal human skin* 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 (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab7800, 1ug/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 haematoxylin 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

  • All lanes :

    Lane 1 : A431 whole cell lysate
    Lane 2 : Human skin whole tissue lysate
    Lane 3 : SH-SY5Y whole cell lysate (negative control)

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Observed band size: 55 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 70 kDa (possible cross reactivity)



    This blot was produced using a 4-12% Bis-tris gel 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 then blocked for an hour using 3% milk before ab7800 and ab181602 (Rabbit anti GAPDH), were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • ab7800 staining Cytokeratin 14 in A431 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% 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 ab7800 at 0.1ugml then detected with an Alexa Fluor® 488 goat anti-mouse secondary antibody (ab150117) at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab202272, Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).

  • ab7800 staining Cytokeratin 14 in Human normal skin tissue sections by IHC-P (Formaldehyde-fixed, Paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with 10% Serum for 30 minutes at 21°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). The sample was incubated with primary antibody (1/100 in PBS + 0.5% Tween-20 + 0.5% BSA)) at 21°C for 30 minutes. An undiluted HRP-conjugated goat polyclonal to mouse IgG was used as secondary antibody.

    See Abreview

  • ab7800 was used to stain mouse prostate.

  • Anti-Cytokeratin 14 antibody [LL002] (ab7800) at 2 µg/ml + Human HaCaT whole cell lysate at 30 µg

    Secondary
    Goat Anti-mouse IgG Polyclonal at 1/20000 dilution

    Developed using the ECL technique.

    Observed band size: 55 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking Step: 5% Milk for 12 hours at 4°C
    Gel Running Conditions: 15%,6V,50min; Reduced; Denaturing

    See Abreview

  • Immunofluorescence analysis of mouse mammary epithelial cells, staining Cytokeratin 14 (red) with ab7800.
  • Immunohistochemical analysis of mouse mammary duct tissue, staining Cytokeratin 14 (red) with ab7800.

    Antigen retrieval was carried out on paraffin-embedded sections by boiling in citrate buffer (pH 6) for 18 minutes in a microwave. Sections were then blocked for 1.5 hours in blocking reagents, before incubating with primary antibody (0.26 µg/ml) overnight at 4°C. An AlexaFluor®555-conjugated goat anti-mouse IgG was used as the secondary antibody

References

This product has been referenced in:
  • Boecker W  et al. Spatial analysis of p63, K5 and K7 defines two groups of progenitor cells that differentially contribute to the maintenance of normal sebaceous glands, extraocular sebaceous carcinoma and benign sebaceous tumors. J Dermatol 46:249-258 (2019). Read more (PubMed: 30663115) »
  • Noguchi S  et al. Beclin 1 regulates recycling endosome and is required for skin development in mice. Commun Biol 2:37 (2019). Read more (PubMed: 30701202) »
See all 105 Publications for this product

Customer reviews and Q&As

1-10 of 25 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (Primary airway epithelial cells)
Permeabilization
No
Specification
Primary airway epithelial cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
95% ethanol

Dr. Egi Kardia

Verified customer

Submitted Sep 10 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Breast (normal and tumoral))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris ph9 / citrate pH6
Permeabilization
Yes - Tween 20 (0.1%)
Specification
Breast (normal and tumoral)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 10 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Salivary gland)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Salivary gland
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 08 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH9
Permeabilization
Yes - 0.05 TX 100
Specification
Skin
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Formaldehyde

Dr. Vincenzo Miragliotta

Verified customer

Submitted Jun 05 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Human prostate cancer cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Human prostate cancer cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 18 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mammary gland)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 1mM EDTA 95ºC pH=8.0
Permeabilization
Yes - PBS with 3% Triton
Specification
Mammary gland
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 04 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Breast -MCF7)
Specification
Breast -MCF7
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 07 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (Breast cancer cell lines)
Specification
Breast cancer cell lines
Permeabilization
Yes - Triton-X100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 11 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
(agent) for 1 hour(s) and 30 minute(s) · Concentration: 2% · Temperature: 25°C
Sample
Human Tissue sections (Human Skin)
Specification
Human Skin
Permeabilization
Yes - 0.1% tritonx-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 09 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric acid
Sample
Mouse Tissue sections (Colon)
Specification
Colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 06 2014

1-10 of 25 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up