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  1. Link

    cytokeratin-14-antibody-sp53-ab119695.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Intermediate Filaments Class I Cytokeratins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cytokeratin 14 antibody [SP53] (ab119695)

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Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Western blot - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Flow Cytometry - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
  • Anti-Cytokeratin 14 antibody [SP53] (ab119695)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP53] to Cytokeratin 14
  • Suitable for: ICC/IF, IHC-P, WB, Flow Cyt, IHC-Fr
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)

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Overview

  • Product name

    Anti-Cytokeratin 14 antibody [SP53]
    See all Cytokeratin 14 primary antibodies
  • Description

    Rabbit monoclonal [SP53] to Cytokeratin 14
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Pig
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431 cell lysate and human skin tissue lysate. IHC-P: Human prostate tissue. IHC-Fr: Mouse and Rat skin tissue. ICC/IF: A431 cells. Flow Cyt: A431 cells.
  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Purification notes

    Purified from TCS by protein A/G.
  • Clonality

    Monoclonal
  • Clone number

    SP53
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class I
    • Cytokeratins

Associated products

  • Alternative Versions

    • Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
  • KO cell lysates

    • Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (ab261706)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab119695 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
ICC/IF (1)
Use a concentration of 5 µg/ml.
IHC-P (1)
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/25. Detects a band of approximately 50 kDa (predicted molecular weight: 52 kDa).
Flow Cyt
Use at an assay dependent concentration.

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

IHC-Fr (1)
Use at an assay dependent concentration.
Notes
ICC/IF
Use a concentration of 5 µg/ml.
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/25. Detects a band of approximately 50 kDa (predicted molecular weight: 52 kDa).
Flow Cyt
Use at an assay dependent concentration.

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

IHC-Fr
Use at an assay dependent concentration.

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.
  • Target information above from: UniProt accession P02533 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 404111 Cow
    • Entrez Gene: 3861 Human
    • Entrez Gene: 16664 Mouse
    • Entrez Gene: 287701 Rat
    • Omim: 148066 Human
    • SwissProt: P05785 Cow
    • SwissProt: P02533 Human
    • SwissProt: Q61781 Mouse
    • SwissProt: Q6IFV1 Rat
    • Unigene: 654380 Human
    • Unigene: 439898 Mouse
    • Unigene: 145931 Rat
    • Unigene: 153972 Rat
    see all
  • 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 [SP53] (ab119695)
    Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)

    Immunohistochemistry (Frozen) analysis of mouse skin tissue section labeling Cytokeratin 14 with purified ab119695. Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    ab119695 staining KRT14 in wild-type A431 cells (top panel) and KRT14 knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then 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 with ab119695 at 5μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Western blot - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Western blot - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    All lanes : Anti-Cytokeratin 14 antibody [SP53] (ab119695) at 1/93 dilution

    Lane 1 : Wild-type A431 whole cell lysate
    Lane 2 : KRT14 knockout A431 whole cell lysate
    Lane 3 : Human skin whole tissue lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab119695 observed at 52 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab119695 was shown to react with KRT14 in A431 wild-type cells in Western blot. Loss of signal was observed when KRT14 knockout sample was used. A431 wild-type and KRT14 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab119695 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 93 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] (ab119695)

    Immunocytochemistry/ Immunofluorescence analysis of A431(human epidermoid carcinoma epithelial cell) cells labeling Cytokeratin 14 with purified ab119695. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Flow Cytometry - Anti-Cytokeratin 14 antibody [SP53] (ab119695)

    Flow cytometry analysis of A431 (human epidermoid carcinoma) labeling Cytokeratin 14 with purified ab119695 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as a secondary antibody. Isotype control -Rabbit monoclonal IgG (ab172730) (Black). Unlableled control -Unlabelled cells (blue).

  • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)

    Immunohistochemistry (Frozen) analysis of rat skin tissue section labeling Cytokeratin 14 with purified ab119695. Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Human prostate tissue stained with ab119695 at a dilution of 1/100.
  • Anti-Cytokeratin 14 antibody [SP53] (ab119695)
    Anti-Cytokeratin 14 antibody [SP53] (ab119695)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (3)

Publishing research using ab119695? Please let us know so that we can cite the reference in this datasheet.

ab119695 has been referenced in 3 publications.

  • Gao H  et al. Characterization of murine mammary stem/progenitor cells in a D-galactose-induced aging model. Aging (Albany NY) 13:11762-11773 (2021). PubMed: 33878032
  • Lin Z  et al. Epidermal stem cells maintain stemness via a biomimetic micro/nanofiber scaffold that promotes wound healing by activating the Notch signaling pathway. Stem Cell Res Ther 12:341 (2021). PubMed: 34112252
  • Su Y  et al. Pre-aggregation of scalp progenitor dermal and epidermal stem cells activates the WNT pathway and promotes hair follicle formation in in vitro and in vivo systems. Stem Cell Res Ther 10:403 (2019). PubMed: 31856904

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Immunocytochemistry abreview for Anti-Cytokeratin 14 antibody [SP53]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cultured Cells (Keratinocytes)
Permeabilization
No
Specification
Keratinocytes
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: RT°C
Fixative
Acetone
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 15 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Cytokeratin 14 antibody [SP53]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Keratinocytes)
Permeabilization
No
Specification
Keratinocytes
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 23°C
Fixative
Acetone
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 02 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 14 antibody [SP53]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Hair follicle)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA
Permeabilization
No
Specification
Hair follicle
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 17 2021

Immunohistochemistry (Frozen sections) abreview for Anti-Cytokeratin 14 antibody [SP53]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 21°C
Fixative
Acetone
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 17 2020

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