Overview

  • Product name

    Human Cytokeratin 14 ELISA Kit
    See all Cytokeratin 14 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Urine 5 4.1%
    Inter-assay
    Sample n Mean SD CV%
    Urine 3 11.1%
  • Sample type

    Saliva, Urine, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    4.9 pg/ml
  • Range

    31.3 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 102 99% - 106%
    Urine 110 108% - 114%
    Cell culture extracts 99 93% - 106%
    Tissue Extracts 91 88% - 94%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse
  • Product overview

    Cytokeratin 14 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Cytokeratin 14 protein in Human urine, saliva, cell and tissue extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Cytokeratin 14 is an intermediate filament protein in the keratin family and is a component of the cytoskeletal scaffold in epithelial cells. Cytokeratin 14 is mutated in epidermolysis bullosa simplex, Naegeli-Franceschetti-Jadassohn syndrome and Dermatopathia pigmentosa reticularis. Cytokeratin 14 is a 472 residue protein and the antibodies in this kit were raised to the C-terminal 357 residues.


    Sensitivity:


    Samples diluted in Sample Diluent NS: 13.8 pg/mL


    Samples diluted in 1X Cell Extraction Buffer PTR: 4.9 pg/mL

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BR 1 x 6ml
    10X Human Cytokeratin 14 Capture Antibody 1 x 600µl
    10X Human Cytokeratin 14 Detector Antibody 1 x 600µl
    Human Cytokeratin 14 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
  • Alternative names

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

Applications

Our Abpromise guarantee covers the use of ab226895 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Cytokeratin 14 were measured in duplicates, interpolated from the Cytokeratin 14 standard curves and corrected for sample dilution. Undiluted samples are as follows: saliva 50% and urine 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytokeratin 14 concentration was determined to be 3.0 ng/mL in saliva and 1.06 ng/mL in urine.

  • The concentrations of Cytokeratin 14 were measured in duplicate and interpolated from the Cytokeratin 14 standard curve and corrected for sample dilution and extract load. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytokeratin 14 concentration was determined to be 396 pg/µg skin extract and 491 pg/µg in HACAT cell extract.

Protocols

References

ab226895 has not yet been referenced specifically in any publications.

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