Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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)

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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