Ki67 Antibody | AbD02531

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Ki67 Antibody | AbD02531 gallery image 1

Staining of formalin-fixed paraffin embedded human tonsil section with human anti Ki-67 (HCA006). A: Overview, B: x400 magnification

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Ki67 Antibody | AbD02531 gallery image 2

Staining of frozen human tonsil section using human anti Ki-67 (HCA006)

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Ki67 Antibody | AbD02531 gallery image 3

Staining of MCF-7 with Human anti Ki-67 (HCA006, green) counterstained with Phalloidin (red) and Hoechst (blue)

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Ki67 Antibody | AbD02531 gallery image 4

Staining of paraffin embedded human lymph node with Human anti Ki-67
A) Immunohistochemistry (paraffin sections) was carried out using the Human anti-Ki67 antibody carrying a His-6 tag (HCA006) at 1/50 on Human Lymph Node LN6 Hodgkins tissue. Antigen retrieval was conducted using citrate buffer (pH6) treatment. For detection, HRP conjugated Mouse anti-Histidine tag (MCA5995P ) was applied at 1/50 (brown). B) As a negative control, Mouse anti-Histidine tag (MCA5995P ) was applied at 1/50 on Human Lymph Node LN6 Hodgkins tissue that had been treated in the same way. Sections were counterstained with Hematoxylin (blue). Images were visualized at x20 zoom

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  • Human anti Ki67
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    AbD02531
  • Isotype
    HuCAL Fab bivalent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    HCA006C, E, IF, P *, WBdatasheet pdfdatasheet pdf0.1 mg
    HCA006
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
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    • Human anti Ki67 antibody, clone AbD02531, recognizes the sequence GFKELFQ, which can be found eight times on the human Ki67 protein: Amino acid positions 1104-1110, 1226-1232, 1348-1354, 1590-1596, 1956-1962, 2078-2084, 2318-2324, and 2439-2445.

      Ki-67 antigen is expressed in proliferating cells but not in quiescent cells. Expression of this antigen occurs preferentially during late G1, S, G2, and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected. Consequently, Ki-67 antigen expression is used in tumor pathology to detect proliferating cells in neoplastic diseases. In cultured cells, Ki-67 is expressed in the nucleolus of interphase cells.
      The Ki-67 gene contains 15 exons. The Ki-67 repeat region, within which there is a 22-amino acid Ki-67 motif, is encoded by exon 13. The shorter isoform lacks exon 7. Northern blot analysis revealed multiple transcripts ranging from approximately 8.9 to 12.5 kb in proliferating but not quiescent cells. Immunoblot analysis showed expression of 320- and 359-kDa proteins. Sequence analysis predicted that the short-lived 2,896- and 3,256-amino acid protein isoforms contain potential nuclear targeting signals, over 200 potential phosphorylation sites, 19 N-myristoylation sites, 3 amidation sites, and numerous PEST sites. Antisense oligonucleotides inhibited cellular proliferation in a dose-dependent manner, suggesting that Ki-67 protein expression may be an absolute requirement for cell proliferation. Clone AbD02531 is also known as clone 2531.
    • Intended Use
    • Target Species
      Human
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Rhesus MonkeyExpected from Sequence
      BovineExpected from Sequence
      ChimpanzeeExpected from Sequence
      DogExpected from Sequence
      MacaqueExpected from Sequence
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Bivalent human recombinant Fab selected from the HuCAL® GOLD phage display library. Expressed in E. coli and purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix motif. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the antibody heavy chain -- Liquid.
    • Reconstitution
    • Preparation
      Metal chelate affinity chromatography
    • Preservative Stabilisers
      0.01% Thiomersal
    • Immunogen
      Peptide derived from the human Ki-67 protein. Sequence GFKELFQTPG, coupled via a C-terminal cysteine to carrier proteins.
    • Activity
      Activity was tested by indirect ELISA: The Ki-67 antigen peptide was coupled to human transferrin carrier protein. The coupled antigen (5 μg/ml) plus unrelated control proteins were immobilized on a microtiter plate. Specific binding was monitored by first adding HCA006 (2 μg/ml), then adding a secondary antibody (goat anti-human F(ab')2 fragment specific, AP conjugate. 1:5000 diluted). A fluorescent signal was created by adding the AP sub-strate Attophos. The signal on the antigen is at least 5-fold above background, whereas the signal on the control antigens is less than 1.5-fold above background.
    • Purity
    • Approx. Protein Concentrations
      Antibody concentration 0.5 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. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      12 months from date of despatch.
    • GO Terms
    • UniProt
    • Entrez Gene
    • Acknowledgements
      Sold under license of U.S. Patents 6,300,064, 6,696,248, 6,708,484, 6,753,136, European Patent 0,859,841 and corresponding patents.
      This antibody was developed by Bio-Rad, a wholly owned subsidiary of Bio-Rad Laboratories Inc., Zeppelinstr. 4, 82178 Puchheim, Germany.
    • Licensed Use
      For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
    • 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
      ELISA2ug/ml
      Immunofluorescence
      Immunohistology - Frozen10ug/ml
      Western Blotting5ug/ml
      Immunohistology - Paraffin(1)10ug/ml
      (1)
      This product requires antigen retrieval using heat treatment prior to staining of paraffin sections.Sodium citrate buffer pH 6.0 is recommended for this purpose.

    • 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 appropriate negative/positive controls.
    • Technical Advice
      Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
      Human tonsil.
    • Immunofluorescence
    • Western Blotting
      HCA006 detects bands of approximately 360kDa and 320kDa in Granta cell lysates
    • Instructions For Use

    Additional Ki67 Antibody Formats

    Formats Clone Applications Sizes available
    Ki67 Antibody : Purified AbD02531 C, E, IF, P *, WB 0.1 mg
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Goat anti Human IgG F(ab')2:Alk. Phos.0500-01000.5 mlE, WB
      0500-0100
      Goat anti Human IgG F(ab')2:HRP0500-00990.5 mlE, WB
      0500-0099
      Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
      MCA5995P
      Mouse anti c-MycMCA22001 mgC, E, F *, P, WB*
      MCA2200

      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              Human tonsil.

              References

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