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  1. Link

    cytokeratin-15-antibody-epr1614y-ab52816.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 15 antibody [EPR1614Y] (ab52816)

  • Datasheet
  • SDS
Reviews (2)Q&A (4)References (40)

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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 (Intracellular) - 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, Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

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Primary
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Alexa Fluor® 488 Anti-Cytokeratin 13 antibody [EPR3671] (ab198584)
Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Primary
Product image
Anti-Cytokeratin 15 antibody [EPR1614Y] - BSA and Azide free (ab239850)

View more associated products

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

    Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Human
  • 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 (intra): 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

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR1614Y
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Cytokeratin 15 antibody [EPR1614Y] (ab194065)
    • Alexa Fluor® 647 Anti-Cytokeratin 15 antibody [EPR1614Y] (ab194066)
    • Alexa Fluor® 555 Anti-Cytokeratin 15 antibody [EPR1614Y] (ab214393)
    • Anti-Cytokeratin 15 antibody [EPR1614Y] - BSA and Azide free (ab239850)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

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

    • Human KRT15 knockout A-431 cell line (ab262484)
  • KO cell lysates

    • Human KRT15 knockout A-431 cell lysate (ab263922)

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.

Application Abreviews Notes
ICC/IF
1/50 - 1/100.
Flow Cyt (Intra)
1/100.

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

WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
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.
Flow Cyt (Intra)
1/100.

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

WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
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.
  • Target information above from: UniProt accession P19012 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3866 Human
    • Entrez Gene: 16665 Mouse
    • Omim: 148030 Human
    • SwissProt: P19012 Human
    • SwissProt: Q61414 Mouse
    • Unigene: 654570 Human
    • Unigene: 440010 Mouse
    • 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 (Intracellular) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
      Flow Cytometry (Intracellular) - 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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (40)

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

    ab52816 has been referenced in 40 publications.

    • Zhong P  et al. Low KRT15 expression is associated with poor prognosis in patients with breast invasive carcinoma. Exp Ther Med 21:305 (2021). PubMed: 33717248
    • Morgun EI  et al. Expression of Protein Kinases RIPK-1 and RIPK-3 in Mouse and Human Hair Follicle. Dokl Biochem Biophys 494:252-255 (2020). PubMed: 33119828
    • Polec A  et al. PML Regulates the Epidermal Differentiation Complex and Skin Morphogenesis during Mouse Embryogenesis. Genes (Basel) 11:N/A (2020). PubMed: 32992884
    • Moses RL  et al. Novel epoxy-tiglianes stimulate skin keratinocyte wound healing responses and re-epithelialization via protein kinase C activation. Biochem Pharmacol 178:114048 (2020). PubMed: 32446889
    • Mertsch S  et al. Decellularized human corneal stromal cell sheet as a novel matrix for ocular surface reconstruction. J Tissue Eng Regen Med 14:1318-1332 (2020). PubMed: 32652796
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-6 of 6 Abreviews or Q&A

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

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

    Abcam user community

    Verified customer

    Submitted Jan 28 2020

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

    Inconclusive
    Abreviews
    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

    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

    Abcam community

    Verified customer

    Asked on Oct 16 2012

    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

    Abcam Scientific Support

    Answered on Oct 16 2012

    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

    Abcam community

    Verified customer

    Asked on Oct 15 2012

    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

    Abcam Scientific Support

    Answered on Oct 15 2012

    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

    Abcam community

    Verified customer

    Asked on Dec 14 2011

    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

    Abcam Scientific Support

    Answered on Dec 14 2011

    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

    Abcam community

    Verified customer

    Asked on Dec 14 2011

    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

    Abcam Scientific Support

    Answered on Dec 14 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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