Cytokeratin 19 Antibody | A53-B/A2

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Cytokeratin 19 Antibody | A53-B/A2 gallery image 1

Staining of paraffin embedded human appendix with Mouse anti Human cytokeratin 19 (5552-9009)

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Western blot analysis of A549 human alveolar adenocarcinoma whole cell lysate probed with Mouse anti Human cytokeratin 19 antibody (5552-9009) followed by HRP conjugated Goat anti Mouse IgG, visualized by chemiluminescence

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Sandwich ELISA analysis of human cytokeratin-19 expression using Mouse anti Human cytokeratin 19 antibody, clone BA19 (MCA2145) as a capture reagent and biotinylated Mouse anti Human cytokeratin 19 antibody, clone A53-B/A2 (5552-9009<) as a detection reagent with purified human cytokeratin 19 (Bio-rad catalog; 34002301) as antigen to create the standard curve. Detection is by HRP conjugated streptavidin and substrate. Microtitre plate is read at O.D. 450 nm on the Bio-Rad iMark Microplate Absorbance Reader (Bio-Rad Catalog 168-1135). Normal serum (green), colorectal cancer serum (orange) and breast cancer serum (red) samples at 1:4 dilution are shown

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Published customer image:
Mouse antiHuman cytokeratin 19 antibody, clone A53-B/A2 used for the identification of CK19 expressing cells and tissues by immunohistochemistry and immunofluorescence.
Image caption:
High-content screening of normal human mammary epithelial cells (hMECs). a Schematic representation of a normal human mammary duct in vivo showing the major cell types: basal cells (red) and luminal cells (blue). b Light micrographs of immunohistochemical staining of K19 and K14 on FFPE sections of normal human breast delineating luminal and basal cells respectively. c Representative image of immunofluorescence staining of primary hMECs acquired using the IN Cell Analyser 2000 (GE Healthcare, Silverwater, NSW, Australia) at 20× magnification. Cells were stained for K14 (red) and K19 (blue) and counterstained with DAPI (white). Areas of the four possible cell phenotypes are indicated (arrows): double negative or K14-/K19-; double positive or K14+/K19+; K14+ (K14+/K19-); and K19+ (K14-/K19+). d Identification of cell phenotype on a single cell basis using digital image analysis. The image analysis protocol defines cells based on nuclear segmentation (first column), then subsequently measures the average intensity of K14 (second column) and K19 (third column) within each cell. The four possible outcomes for K14 and K19 are shown by row. e Intensity distributions for K14 (left panels) and K19 (right panels) in primary hMEC culture of negative secondary only control (black) and stained samples plotting their respective cytokeratin distribution (K14, red; K19, blue). f Subpopulation frequency of the four cell phenotypes in the first passage of hMECs for five donors. Mean subpopulation frequencies across (n?=?5–6) technical replicates were calculated as the average percentage of total cell number within each well for each of the five individual donors (± standard error of the mean (SEM)). g Subpopulation frequency with passage and enrichment using FACS (Fluorescence-activated cell sorting). Cells from reduction mammoplasties were dissociated and cultured in vitro (P1). After 5–7 days, cells were harvested and sorted based on expression of CD10 and MUC1 using FACS. CD10+, MUC1+ and unsorted (US) cells were cultured for 5 days (P2) before fixation and analysis of population heterogeneity. Mean subpopulation frequencies were calculated as the average percentage of total cell number within each well for each sample (± SEM). DAPI 4',6-diamidino-2-phenylindole, FACS fluorescence-activated cell sorting, K14 cytokeratin 14, K19 cytokeratin 19, P1 passage 1, P2 passage 2

From: Johnston RL, Wockner L, McCart Reed AE, Wiegmans A, Chenevix-Trench G, Khanna KK, Lakhani SR, Smart CE.
High content screening application for cell-type specific behaviour in heterogeneous primary breast epithelial subpopulations.
Breast Cancer Res. 2016 Feb 9;18(1):18.

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Cytokeratin 19 Antibody | A53-B/A2 gallery image 5

Published customer image:
Mouse antiHuman cytokeratin 19 antibody, clone A53-B/A2 used for the identification of CK19 expressing cells and tissues by immunofluorescence.
Image caption:
Assessment of alternative markers to delineate hMEC subpopulations using high-content screening. a Immunofluorescence staining of hMECs with DAPI (white) and antibodies against K14 (red), K19 (blue) and K8/18 (green; top panel) or K5 (green; bottom panel). Images acquired at 20× magnification using INCA. b Proportional Venn diagrams depict the mean overall proportions and overlap of K14, K8/18 and K19 expression (top) and K14, K5 and K19 expression (bottom) in first-passage hMEC cultures for donor Q626. c IHC staining of K5 and K8/18 on FFPE sections of normal human breast. Staining for K5 (top left, 10×) delineates both basal cells (i; top right, 20×) and luminal cells (ii; bottom left, 20×). Staining for K8/18 (bottom right, 10×) preferentially delineates basal cells. DAPI 4',6-diamidino-2-phenylindole, K5 cytokeratin 5, K8/18 cytokeratin 8/18, K14 cytokeratin 14, K19 cytokeratin 19

From: Johnston RL, Wockner L, McCart Reed AE, Wiegmans A, Chenevix-Trench G, Khanna KK, Lakhani SR, Smart CE.
High content screening application for cell-type specific behaviour in heterogeneous primary breast epithelial subpopulations.
Breast Cancer Res. 2016 Feb 9;18(1):18.

Enlarge
Cytokeratin 19 Antibody | A53-B/A2 gallery image 6

Published customer image:
Mouse antiHuman cytokeratin 19 antibody, clone A53-B/A2 used for the identification of CK19 expressing cells and tissues by immunofluorescence.
Image caption:
Assessment of alternative markers to delineate hMEC subpopulations using high-content screening. a Immunofluorescence staining of hMECs with DAPI (white) and antibodies against K14 (red), K19 (blue) and K8/18 (green; top panel) or K5 (green; bottom panel). Images acquired at 20× magnification using INCA. b Proportional Venn diagrams depict the mean overall proportions and overlap of K14, K8/18 and K19 expression (top) and K14, K5 and K19 expression (bottom) in first-passage hMEC cultures for donor Q626. c IHC staining of K5 and K8/18 on FFPE sections of normal human breast. Staining for K5 (top left, 10×) delineates both basal cells (i; top right, 20×) and luminal cells (ii; bottom left, 20×). Staining for K8/18 (bottom right, 10×) preferentially delineates basal cells. DAPI 4',6-diamidino-2-phenylindole, K5 cytokeratin 5, K8/18 cytokeratin 8/18, K14 cytokeratin 14, K19 cytokeratin 19

From: Johnston RL, Wockner L, McCart Reed AE, Wiegmans A, Chenevix-Trench G, Khanna KK, Lakhani SR, Smart CE.
High content screening application for cell-type specific behaviour in heterogeneous primary breast epithelial subpopulations.
Breast Cancer Res. 2016 Feb 9;18(1):18.

Enlarge
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  • Mouse anti Human Cytokeratin 19
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    A53-B/A2
  • Isotype
    IgG2a
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    5552-9009E, IF, P *, WBdatasheet pdfdatasheet pdf0.2 mg
    5552-9009
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Human cytokeratin 19 antibody, clone A53-B/A2 recognizes the rod domain (aa 312-335) of human cytokeratin 19 (Böttger; et al. 1995) also known as keratin 19 encoded by the KRT19 gene. Cytokeratin 19 is a 400 amino acid intermediate filament protein lacking a C-terminal tail domain, in contrast to all other intermediate filament proteins.

      Cytokeratin 19 expression is observed in striated muscle where it is involved in forming the association between the contractile apparatus and dystrophin (Stone et al. 2005). Expression is also seen in many ductal and glandular cells together with a restricted set of normal and neoplasic epithelial cells.

      Mouse anti Human cytokeratin 19 antibody, clone A53-B/A2 detects a band of ~45 kDA in Western blotting using an A-549 human alveolar adenocarcinoma cell line lysate.
    • Intended Use
    • Target Species
      Human
    • Product Form
      Purified IgG - liquid
    • Reconstitution
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
    • Preservative Stabilisers
      0.09%Sodium Azide (NaN3)
    • Immunogen
      Human breast cancer cell line MCF-7.
    • Purity
    • Approx. Protein Concentrations
      1.0 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody.
    • Shelf Life
      18 months from date of despatch.
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Immunofluorescence
      Western Blotting
      Immunohistology - Paraffin(1)1/1001/200
      (1)
      This product requires protein digestion pre-treatment of paraffin sections e.g. trypsin or pronase.

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using the appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
      Appendix
    • Immunofluorescence
    • Western Blotting
      In Western blotting 40 kD and 19 kD bands are observed.
    • Instructions For Use

    Additional Cytokeratin 19 Antibody Formats

    Formats Clone Applications Sizes available
    Cytokeratin 19 Antibody : Purified A53-B/A2 E, IF, P *, WB 0.2 mg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              Appendix

              References

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