Recombinant
RabMAb

Anti-Cytokeratin 19 antibody [EPR1579Y] (ab76539)

Overview

  • Product name
    Anti-Cytokeratin 19 antibody [EPR1579Y]
    See all Cytokeratin 19 primary antibodies
  • Description
    Rabbit monoclonal [EPR1579Y] to Cytokeratin 19
  • Host species
    Rabbit
  • Specificity
    In our WB testing this antibody did not recognize mouse Cytokeratin 19, but customers have reported the antibody recognizes the protein in IHC-P with mouse tissue. Thus, we have removed mouse as a "does not react with" species and welcome further feedback from other researchers using this antibody in mouse.
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    within Human Cytokeratin 19 (N terminal). The exact sequence is proprietary.

  • Positive control
    • WB: T47D, HepG2 or SKBR-3 cell lysate; IHC-P: Human stomach adenocarcinoma tissue. IF/ICC: HCT116 cell line.
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.5% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR1579Y
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab76539 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
WB 1/10000 - 1/50000. Predicted molecular weight: 44 kDa.
IP 1/20.
Flow Cyt 1/20.

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

 

IHC-P 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

ICC/IF 1/100 - 1/250.

Target

  • Function
    Involved in the organization of myofibers. Together with KRT8, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.
  • Tissue specificity
    Expressed in a defined zone of basal keratinocytes in the deep outer root sheath of hair follicles. Also observed in sweat gland and mammary gland ductal and secretory cells, bile ducts, gastrointestinal tract, bladder urothelium, oral epithelia, esophagus, ectocervical epithelium (at protein level). Expressed in epidermal basal cells, in nipple epidermis and a defined region of the hair follicle. Also seen in a subset of vascular wall cells in both the veins and artery of human umbilical cord, and in umbilical cord vascular smooth muscle. Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma in structures that contain dystrophin and spectrin.
  • Sequence similarities
    Belongs to the intermediate filament family.
  • Developmental stage
    Present in hair follicles at all stages of development.
  • Domain
    This keratin differs from all other IF proteins in lacking the C-terminal tail domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • 40 kDa keratin intermediate filament antibody
    • CK 19 antibody
    • CK-19 antibody
    • CK19 antibody
    • Cytokeratin 19 antibody
    • Cytokeratin-19 antibody
    • K19 antibody
    • K1C19_HUMAN antibody
    • K1CS antibody
    • Keratin 19 antibody
    • Keratin type I 40 kD antibody
    • Keratin type I 40kD antibody
    • Keratin type I cytoskeletal 19 antibody
    • Keratin, type I cytoskeletal 19 antibody
    • Keratin, type I, 40 kd antibody
    • Keratin-19 antibody
    • KRT19 antibody
    • MGC15366 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling Cytokeratin 19 with Purified ab76539 at 1:1000 dilution (0.12 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
  • ab76539 (purified) at 1:20 dilution (2ug) immunoprecipitating Cytokeratin 19 in SKBR-3 whole cell lysate.
    Lane 1 (input): SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg
    Lane 2 (+): ab76539 & SKBR-3 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76539 in SKBR-3 whole cell lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cytokeratin 19 with purified ab76539 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunofluorescence staining of MCF-7 cells with purified ab76539 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 100% methanol. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

  • All lanes : Anti-Cytokeratin 19 antibody [EPR1579Y] (ab76539) at 1/10000 dilution (unpurified)

    Lane 1 : T47D cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : SKBR-3 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 44 kDa

  • Immunohistochemical staining of human stomach adenocarcinoma tissue with ab76539 (unpurified) at1/100-1/250.

  • ab76539 (unpurified) stained HCT116 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76539 at 1/200 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab76539 (unpurified). Green- CK19 red-PI.

  • Overlay histogram showing MCF7 cells stained with unpurified ab76539 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 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 (ab76539, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

References

This product has been referenced in:
  • Maidji E  et al. Replication of CMV in the gut of HIV-infected individuals and epithelial barrier dysfunction. PLoS Pathog 13:e1006202 (2017). Read more (PubMed: 28241080) »
  • Yanagi Y  et al. In vivo and ex vivo methods of growing a liver bud through tissue connection. Sci Rep 7:14085 (2017). IHC ; Rat . Read more (PubMed: 29074999) »

See all 5 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer Cell lines)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
Cancer Cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted May 02 2018

Although mouse samples were not recommended based on the western blot results below with these antibodies, there is still the possibility that the negative results in mouse might be due to the limited sample types, especially for the antibodies with hi...

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Thank you very much for contacting us. Following your request, I have searched our catalog for all antibodies that match the mentioned criteria, i.e. that recognise human targets and that do not react with mouse. Please find attached a list of ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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