Recombinant
RabMAb

Recombinant Anti-Cytokeratin 19 antibody [EPR1579Y] - BSA and Azide free (ab232566)

Overview

  • Product name

    Anti-Cytokeratin 19 antibody [EPR1579Y] - BSA and Azide free
    See all Cytokeratin 19 primary antibodies
  • Description

    Rabbit monoclonal [EPR1579Y] to Cytokeratin 19 - BSA and Azide free
  • 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: IHC-P, WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • IHC-P: Human lung cancer tissue.
  • General notes

    Ab232566 is the carrier-free version of ab76539. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab232566 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab232566 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
IHC-P Use at an assay dependent concentration.

See IHC antigen retrieval protocols.

WB Use at an assay dependent concentration. Predicted molecular weight: 44 kDa.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

  • 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 Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76539).

References

ab232566 has not yet been referenced specifically in any publications.

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