Overview

  • Product name
    Anti-Cytokeratin 14 antibody [RCK107]
    See all Cytokeratin 14 primary antibodies
  • Description
    Mouse monoclonal [RCK107] to Cytokeratin 14
  • Host species
    Mouse
  • Specificity
    This antibody reacts exclusively with cytokeratin 14 which is present in basal cell compartments of stratified and combined epithelia.
  • Tested applications
    Suitable for: Dot blot, ICC, IHC-Fr, WB, ICC/IF, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Rat, Dog, Human, Pig
  • Immunogen

    Cytoskeletal preparation of TR146 epithelial cells.

  • Positive control
    • cytoskeletal preparation of TR146 epithelial cells, skin tissue lysate (Human)
  • General notes


    Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in human epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 – 7.8. The individual human cytokeratins are designated 1 to 20. The various epithelia in the human body usually express cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. The cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Primary antibody notes
    Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in human epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 – 7.8. The individual human cytokeratins are designated 1 to 20. The various epithelia in the human body usually express cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. The cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays.
  • Clonality
    Monoclonal
  • Clone number
    RCK107
  • Myeloma
    Sp2/0-Ag14
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9220 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
Dot blot 1/100 - 1/1000.
ICC Use at an assay dependent concentration.
IHC-Fr 1/100 - 1/200.
WB 1/100 - 1/1000.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/50 - 1/7000. PubMed: 19812262

Heat retrieval (PubMed 19812262) and Proteinase K ()

Flow Cyt 1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

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

  • Immunohistochemical analysis of swine skin tissue labeling Cytokeratin 14 with ab9220. Showing staining of basal cells.

  • ICC/IF image of ab9220 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9220, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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 1.43µM.

  • Anti-Cytokeratin 14 antibody [RCK107] (ab9220) at 4 µg/ml + Human skin tissue lysate - total protein (ab30166) at 20 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 58 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes
  • Immunohistochemical analysis of dog skin epidermis tissue labelling Cytokeratin 14 using ab9220. Staining of basal keratinocyte layer is observed.

  • Immunohistochemical analysis of dog skin appendages labelling Cytokeratin 14 with ab9220.

References

This product has been referenced in:
  • Zubeldia-Plazaola A  et al. Glucocorticoids promote transition of ductal carcinoma in situ to invasive ductal carcinoma by inducing myoepithelial cell apoptosis. Breast Cancer Res 20:65 (2018). Read more (PubMed: 29973218) »
  • Roth M  et al. Broncho Vaxom (OM-85) modulates rhinovirus docking proteins on human airway epithelial cells via Erk1/2 mitogen activated protein kinase and cAMP. PLoS One 12:e0188010 (2017). Read more (PubMed: 29182620) »
See all 17 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Mouse Tissue sections (skin)
Specification
skin
Permeabilization
Yes - tritonx-100
Fixative
Paraformaldehyde

Ms. Megan Scott

Verified customer

Submitted Aug 27 2013

Answer

Gracias por contactarnos.

Como bien dices, tenemos varios anticuerpos contra los marcadores especificados, sin embargo, que se hayan testado y validado (y por tanto, estén cubiertos por la garantía Abpromise) en inmunocitoquímica son muchos menos.

La citoqueratina 14 es en efecto un buen marcador de células miopiteliales en glándula mamaria, y aunque ninguno de los tres anticuerpos testados en ICC se haya probado en estas células, todos ellos deberían funcionar.

Los enlaces a las datasheets son:

https://www.abcam.com/Cytokeratin-14-antibody-RCK107-ab9220.html

https://www.abcam.com/Cytokeratin-14-antibody-EPR1612-ab108417.html

https://www.abcam.com/Cytokeratin-14-antibody-SPK-14-ab9384.html

Cualquiera de ellos es una buena opción. El primero tiene una imagen de células HepG2 teñidas con el anticuerpo. Los rabbit monoclonals (ab108414) suelen ser muy limpios y específicos. De todas maneras todos ellos están garantizados en ICC, por lo que en caso de ni funcionar correctamente se os mandaría un reemplazo (del mismo o diferente anticuerpo) gratuito o un reembolso del importe.

Otros marcadores de celular mioepiteliales son la citoqueratina 17, p-cadherin o, Vimentin. Para este último tenemos varios anticuerpos. Para vuestro caso concreto ab8978 puede ser una buena opción, y como podéis observar en la pestaña “Abreviews” otros usuarios lo han utilizado obteniendo excelentes resultados:

https://www.abcam.com/Vimentin-antibody-RV202-Neural-Stem-Cell-Marker-ab8978.html

Contra la citoquina 19 las alternativas son mayores, y aunque se hayan usado en líneas celulares muy distintas, podéis ver los Reviews de otros usuarios, que vuelven a ser muy positivos en ICC.

https://www.abcam.com/Cytokeratin-19-antibody-ab15463.html

https://www.abcam.com/Cytokeratin-19-antibody-RCK108-ab9221.html

En cualquier caso como mencioné antes, todos tienen una garantía de 6 meses, y en el caso de no funcionar correctamente se os mandaría un reemplazo o rembolso.

Actualmente tenemos una promoción por la que por la compra de 3 productos se os regala uno adicional (un 4x3). Si necesitáis más información sobre alguno de estos productos, un presupuesto o alguna aclaración, por favor, no dudes en contactarme de nuevo.

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Answer

Thank you for your email and the feedback. I am glad to hear that some of our antibodies work very well for you. As for the 2 antibodies you mentioned that did not work as expected (ab38890 and ab9220), could you please provide a few more protocol details as well as order or PO numbers so that we can resolve these issues according to our Abpromise guarantee (i.e. free of charge replacement, credit or refund if purchased within the last 6 months)? What tissues, type of antigen retrieval, antibody dilutions, blocking conditions and detection systems were used for each antibody? I look forward to hear back from you and to assist you in resolving these problems to your satisfaction. 

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Acetone
Permeabilization
No
Blocking step
Casein as blocking agent for 10 minute(s) · Concentration: 0.25µg/mL · Temperature: 20°C

Dr. Vaughan Feisst

Verified customer

Submitted Jul 20 2011

Answer

I'm sorry to hear you are having a problem with ab9220 and ab18389. I was able to find out that the antibody ab18389 has been used in a paper by Ishida-Yamamoto A et al. 1991. Invest Derm. 97 (6); 959 where a detailed protocol is described, unfortunately I was unable to access this paper but I hope this can help you. I'm also trying to find the dilutions used for this antibody, I can confirm that ab9220 has been tested on frozen sections fixed with ice cold methanol/acetone for 5 min and was used neat or at 1:10 dilution (as it is unpurified tissue culture supernatant), is this the dilution range you tested? There are also several publications which have used ab9220: Vos JH et al. The expression of keratins, vimentin, neurofilament proteins, smooth muscle actin, neuron-specific enolase, and synaptophysin in tumors of the specific glands in the canine anal region. Vet Pathol 30:352-61 (1993). PubMed: 8212457 Vos JH et al. Immunohistochemistry with keratin monoclonal antibodies in canine tissues: urogenital tract, respiratory tract, (neuro-)endocrine tissues, choroid plexus and spinal cord. Zentralbl Veterinarmed A 39:721-40 (1992). PubMed: 1283472 Wetzels RH et al. Basal cell-specific and hyperproliferation-related keratins in human breast cancer. Am J Pathol 138:751-63 (1991). PubMed: 1705754 I hope this information will help you and will let you know the dilution used for ab18389 as soon as I find out,

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