Anti-Cytokeratin 14 antibody [RCK107] (ab9220)

Reviews (2)Q&A (3)References (21)

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

    • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)

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

    • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      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.

    • Western blot - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      Western blot - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      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
    • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)

      Immunohistochemical analysis of dog skin epidermis tissue labelling Cytokeratin 14 using ab9220. Staining of basal keratinocyte layer is observed.

    • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)
      Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [RCK107] (ab9220)

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

    References

    This product has been referenced in:

    • Shrestha R  et al. Aberrant hiPSCs-Derived from Human Keratinocytes Differentiates into 3D Retinal Organoids that Acquire Mature Photoreceptors. Cells 8:N/A (2019). Read more (PubMed: 30634512) »
    • 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) »
    See all 21 Publications for this product

    Publishing research using ab9220? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-Cytokeratin 14 antibody [RCK107]

     Poor
    Abreviews
    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
    Read More

    Ms. Megan Scott

    Verified customer

    Submitted Aug 27 2013

    Question

    Hola,
    estoy interesada en comprar anticuerpos anti-citoqueratina que me
    permitan distinguir células mioepiteliales de células luminales de la
    glándula mamaria humana. Serían las citoqueratina 14 para marcar las
    mioepiteliales y citokeratina 19 para las luminales. He visto que tienen
    una gran oferta de citoqueratinas, buscaba su consejo sobre cual en
    concreto comprar para marcar estas células en particular.
    Además como segundo marcador necesito una vimentina para marcar también
    las mioepiteliales, ¿cuál me recomiendan?
    Gracias

    Read More
    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.

    Read More

    Question

    The antibody that I mentioned having problems with was ab38890 rb pcAb to BrdU. We wanted to use it with sections of bovine mammary gland just to give us another option for dual or triple staining. We routinely use a mouse monoclonal antibody from Roche (clone BMC9318) in our assays, so I had some idea of the expected proliferation index. When all of the nuclei stained, we determined that it would not be usable for our purposes. I did check the ab reviews and noticed that another researcher had the same problem. Other antibodies from Abcam that we use routinely and really like are: ab5694 rb pcAb to alpha smooth muscle actin ab16667 rb mAb to Ki-67 clone SP6 (we can do dual staining with Ki-67 and BrdU) Because we routinely do IHC in formalin-fixed, paraffin embedded bovine tissue, I especially like the information on your website about species cross-reactivity and applications. We sometimes try antibodies just to see if they work even if your website doesn't list cow and IHC-P. Most recently I used ab9220, a mouse mAb to CK14. I was hoping that it would recognize CK14 in . It did not--even with antigen retrieval. I do recommend your website often because of the information you provide there. I noticed recently that you have added a PDF datasheet option.

    Read More
    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. 

    Read More

    Immunohistochemistry (Frozen sections) abreview for Anti-Cytokeratin 14 antibody [RCK107]

     Good
    Abreviews
    abreview image
    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
    Read More

    Dr. Vaughan Feisst

    Verified customer

    Submitted Jul 20 2011

    Question

    BATCH NUMBER 99496 ORDER NUMBER 41493 DESCRIPTION OF THE PROBLEM No staining SAMPLE frozen sections of gingiva, palate and bronchial biopsies PRIMARY ANTIBODY overnight at 4 C or overnight RT DETECTION METHOD AEC POSITIVE AND NEGATIVE CONTROLS USED negative without primary antibody ANTIBODY STORAGE CONDITIONS +4 C FIXATION OF SAMPLE acetone 60% /MeOH 40% or 100% acetone BLOCKING CONDITIONS peroxydase blocking reagent of Dako 20 minutes room temp(RT). protein block. of Dako 20 minutes RT SECONDARY ANTIBODY En Vision (labelled polymer for mouse)of Dako 2 hours rt or anti mouse 1/100 diluted in antibody diluting buffer of Dako 1 hour rt + ABC HRP 1/100 diluted in peroxydase stabiliting buffer of Dako 1 hour rt HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 63 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES I bought also your CK-1 ab18389 and it's the same thing.

    Read More
    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,

    Read More

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