Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)

Reviews (2)Q&A (4)References (27)
Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Flow Cytometry - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • OI-RD Scanning - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
  • Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1614Y] to Cytokeratin 15
  • Suitable for: ICC/IF, WB, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Cytokeratin 15 antibody [EPR1614Y]
    See all Cytokeratin 15 primary antibodies

  • Description

    Rabbit monoclonal [EPR1614Y] to Cytokeratin 15

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Cytokeratin 15 aa 400-500 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Mouse skin tissue lysates; A-431 cell lysate. IHC-P: Human squamous cervical carcinoma, normal skin, cervical carcinoma tissue ICC/IF: HeLa cells. Flow cyt: A-431 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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. Stable for 12 months at -20°C.

  • Dissociation constant (KD)

    KD = 5.60 x 10 -10 M
    Learn more about KD

  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    EPR1614Y

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52816 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Mouse
Application Abreviews Notes
ICC/IF
1/50 - 1/100.
WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
Flow Cyt
1/100.

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

 
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
ICC/IF
1/50 - 1/100.
WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
Flow Cyt
1/100.

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

 
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Tissue specificity

      Expressed in a discontinuous manner in the basal cell layer of adult skin epidermis, but continuously in the basal layer of fetal skin epidermis and nail. Also expressed in the outer root sheath above the hair bulb in hair follicle (at protein level). Expressed homogeneously in all cell layers of the esophagus and exocervix, but detected in the basal cell layer only of oral mucosa, skin and in the basal plus the next two layers of the suprabasal epithelium of the palate.

    • Sequence similarities

      Belongs to the intermediate filament family.
    • Information by UniProt

    • Database links

      • Alternative names

        • AI528832 antibody
        • CK 15 antibody
        • CK-15 antibody
        • CK15 antibody
        • Cytokeratin-15 antibody
        • Cytokeratin15 antibody
        • K15 antibody
        • K1C15_HUMAN antibody
        • K1CO antibody
        • Ka15 antibody
        • Keratin 15 antibody
        • Keratin 15 basic antibody
        • Keratin 15 beta antibody
        • Keratin antibody
        • Keratin complex 1 acidic gene 15 antibody
        • Keratin type I cytoskeletal 15 antibody
        • Keratin-15 antibody
        • Keratin15 antibody
        • KRT 15 antibody
        • Krt1-15 antibody
        • KRT15 antibody
        • KRTB antibody
        • KRTL15 antibody
        • Type I cytoskeletal 15 antibody
        • Type I keratin Ka15 antibody
        see all

      Images

      • Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        All lanes : Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816) at 1/10000 dilution

        Lane 1 : Wild-type A431 cell lysate
        Lane 2 : KRT15 knockout A431 cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 45 kDa
        Observed band size: 50 kDa
        why is the actual band size different from the predicted?



        Lanes 1 - 2: Merged signal (red and green). Green - ab52816 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

        ab52816 was shown to react with Cytokeratin 15 in wild-type A431 cells in Western blot with loss of signal observed in KRT15 knockout cell line ab263922 (knockout cell lysate ab262484). Wild-type A431 and KRT15 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab52816 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 h at room temperature before imaging.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)

        ab52816 at 1/100 dilution staining Cytokeratin 15 in human squamous cervical carcinoma by Immunohistochemistry, Paraffin embedded tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816) at 1/100000 dilution + Mouse skin tissue lysates at 20 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

        Predicted band size: 45 kDa
        Observed band size: 45 kDa



        Blocking buffer and concentration: 5% NFDM/TBST
        Diluting buffer and concentration: 5% NFDM /TBST

        Exposure time: 5.5 seconds

      • Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Western blot - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816) at 1/10000 dilution + fetal thymus lysate at 10 µg

        Secondary
        goat anti-rabbit HRP at 1/2000 dilution

        Predicted band size: 45 kDa
        Observed band size: 45 kDa

      • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        ab52816 at 1/50 dilution staining Cytokeratin 15 in HeLa cells by Immunocytochemistry.
      • Flow Cytometry - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Flow Cytometry - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Overlay histogram showing A431 cells stained with ab52816 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52816,1/100 dilution ) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was was rabbit IgG (monoclonal) ( 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)

        Fluorescent immunohistochemical analysis of paraffin-embedded human normal skin tissue using ab52816. Green-CK15 red-PI

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)

        Fluorescent immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue using ab52816. Green-CK15 red-PI.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • OI-RD Scanning - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        OI-RD Scanning - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD
      • Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
        Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (27)

      Publishing research using ab52816? Please let us know so that we can cite the reference in this datasheet.

      ab52816 has been referenced in 27 publications.

      • Aldehlawi H  et al. The monoclonal antibody EPR1614Y against the stem cell biomarker keratin K15 lacks specificity and reacts with other keratins. Sci Rep 9:1943 (2019). PubMed: 30760780
      • Kim YH  et al. Induction of cardiomyocyte-like cells from hair follicle cells in mice. Int J Mol Med 43:2230-2240 (2019). PubMed: 30864673
      • Rostampour N  et al. Expression of new genes in vertebrate tooth development and p63 signaling. Dev Dyn N/A:N/A (2019). PubMed: 30875130
      • Chu SY  et al. Mechanical stretch induces hair regeneration through the alternative activation of macrophages. Nat Commun 10:1524 (2019). PubMed: 30944305
      • Gouveia RM  et al. YAP, ?Np63, and ß-Catenin Signaling Pathways Are Involved in the Modulation of Corneal Epithelial Stem Cell Phenotype Induced by Substrate Stiffness. Cells 8:N/A (2019). PubMed: 31013745
      View all Publications for this product

      Customer reviews and Q&As

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      1-6 of 6 Abreviews or Q&A

      IHC - Wholemount abreview for Anti-Cytokeratin 15 antibody [EPR1614Y]

       Excellent
      Abreviews
      abreview image
      Application
      IHC - Wholemount
      Sample
      Mouse Tissue (Cornea)
      Specification
      Cornea
      Read More

      Abcam user community

      Verified customer

      Submitted Jan 28 2020

      Question

      Thanks for your e.mail .. I am asking .. can I used them immunofluorescence (IF) for paraffin embedding section ..

      Do you have protocol start with ..

      Many thanks

      Read More
      Answer

      Thank you for your reply.

      I have to confirm that we did not test this antibody in IF with paraffin embedded sections.

      This is in theory possible. Although often high background occurs with this method.

      Please follow this link for a protocol:

      https://www.abcam.com/index.html?pageconfig=resource&rid=11459

      I can also recommend this page for in depth information about IHC:

      http://www.ihcworld.com/knowledge-center.htm

      I hope this information is helpful and wish you good luck with your research.

      Read More

      Question

      Dear Sir/ Madam

      I ordered the following antibodies:

      1) anti-cytokeratin 15 antibodies9(EPR1614Y).(AB52816)



      2) anti-BCRP/ABCG2 antibody (BXP-53).(ab24115)



      I am wondering whether those antibodies can b used in IF , paraffin section / in what concentration .



      many thanks

      Read More
      Answer

      Thank you for your inquiry.

      I am happy to confirm that both antibodies are tested and guarantee for IHC-P.

      1.) We used ab52816 successfully in a dilution of 1/100 in IHC-P with human squamous cervical carcinoma by Immunohistochemistry, Paraffin embedded tissue.

      2.) ab24115 was used at a dilution od 1/400 in the following publication:

      http://www.ncbi.nlm.nih.gov/pubmed/20472681?dopt=Abstract

      Please not that these dilution factors are only recommended starting points for your own optimization experiments.

      I wish you good luck with your experiments.

      Read More

      Question

      Dear Abcam,   I purchased a Rabbit monoclonal [EPR1614Y] to cytokeratin 15 (ab52816) four months ago. I aliquoted smaller volumes and stored at -20C as per your data sheet and kept some at 4C. When the aliquot at 4C finished I started using the ones from -20 and since then I am not getting any band of K15. I have used high concentrations as well (1:1000) and use 5% milk. Can you please suggest what may have gone wrong and if possible send me a new aliquot of the antibody?   Many thanks,

      Read More
      Answer

      Thank you for again confirming these details and for your cooperation.   As requested, I have issued a free of charge replacement with the order number xxxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

      Read More

      Question

      Dear Abcam,   Thank you for replying.   I do not have the order or product number but I bought it this summer. I use standard western blot procedure using nitrocellulose membrane, I use 5% milk for blocking for 15 mins and also 5% milk for primary and secondary antibody for 1 hr at room temperature. I have always used this protocol and never had any problems including the antibody I am complaining about. I have increased the concentration of primary and secondary to 1:1000 and exposed for 2 mins whereas I normally get a strong band even after 30 sec of exposure. I am using the lysates of immortalised keratinocytes NEB1, KEB11 and HaCaT.   It will be of great help if you can suggest anything or provide a new sample of the antibody.   Many thanks

      Read More
      Answer

      Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused.  I would like to offer you a free of charge replacement for ab52816. I have found an order placed xxxxxx, number xxxx. This was shipped to xxxxx. Is this correct? As soon as we can confirm this information I can send you a free of charge replacement antibody. I wish you the best of luck with your research.  

      Read More

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Cytokeratin 15 antibody [EPR1614Y]

       Inconclusive
      Abreviews
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (skin section)
      Specification
      skin section
      Fixative
      Methanol
      Permeabilization
      Yes - acetone
      Blocking step
      BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: rt°C
      Read More

      Ms. Cinzia Chinnici

      Verified customer

      Submitted Aug 20 2009

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