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    alexa-fluor-488-ki67-antibody-epr3610-ab197234.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (2) Submit a question References (12)

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Immunocytochemistry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
  • Immunocytochemistry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
  • Flow Cytometry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
  • Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 488 Rabbit monoclonal [EPR3610] to Ki67
  • Suitable for: Flow Cyt, ICC
  • Knockout validated
  • Reacts with: Human
  • Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm

You may also be interested in

Primary
Product image
Alexa Fluor® 647 Anti-Ki67 antibody [EPR3610] (ab196907)

View more associated products

Overview

  • Product name

    Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610]
    See all Ki67 primary antibodies
  • Description

    Alexa Fluor® 488 Rabbit monoclonal [EPR3610] to Ki67
  • Host species

    Rabbit
  • Conjugation

    Alexa Fluor® 488. Ex: 495nm, Em: 519nm
  • Tested applications

    Suitable for: Flow Cyt, ICCmore details
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human Ki67 aa 1050-1150. The exact sequence is proprietary.

  • Positive control

    • ICC: HeLa cells, HAP1 WT and HAP1-Ki67 KO. Flow Cyt: HeLa cells.
  • General notes

    Alternative versions available:
    Anti-Ki67 antibody [EPR3610] (ab92742) - Knockout validated

    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.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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. Avoid freeze / thaw cycle. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3610
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Markers
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Replication
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Cancer
    • Cell cycle
    • Cell division
    • Cancer
    • Tumor biomarkers
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Ki67 antibody [EPR3610] (ab196907)
    • Anti-Ki67 antibody [EPR3610] - BSA and Azide free (ab209897)
    • Alexa Fluor® 568 Anti-Ki67 antibody [EPR3610] (ab211968)
    • HRP Anti-Ki67 antibody [EPR3610] (ab212215)
    • Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610] (ab215226)
    • Alexa Fluor® 594 Anti-Ki67 antibody [EPR3610] (ab216709)
    • Anti-Ki67 antibody [EPR3610] (ab92742)
  • Isotype control

    • Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199091)

Applications

Our Abpromise guarantee covers the use of ab197234 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
Flow Cyt 1/50.
ICC 1/100.

This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 - 20 min) and 100% methanol (5 min).

It is recommended to incubate cells with 0.1% Triton-X for 5 min to detect nuclear antigen.

Positive Control: HeLa and HAP1 cells / Rt cardiomyocytes

Target

  • Function

    Required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly (PubMed:27362226). Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface (PubMed:27362226). Prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier: the protein has a high net electrical charge and acts as a surfactant, dispersing chromosomes and enabling independent chromosome motility (PubMed:27362226). Binds DNA, with a preference for supercoiled DNA and AT-rich DNA (PubMed:10878551). Does not contribute to the internal structure of mitotic chromosomes (By similarity). May play a role in chromatin organization (PubMed:24867636). It is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in maintaining mitotic chromosomes dispersed.
  • Sequence similarities

    Contains 1 FHA domain.
    Contains 16 K167R repeats.
    Contains 1 PP1-binding domain.
  • Developmental stage

    Expression 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 (at protein level) (PubMed:6206131). Present at highest level in G2 phase and during mitosis (at protein level). In interphase, forms fiber-like structures in fibrillarin-deficient regions surrounding nucleoli (PubMed:2674163, PubMed:8799815).
  • Post-translational
    modifications

    Phosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA.
  • Cellular localization

    Chromosome. Nucleus. Nucleus, nucleolus. Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the mitotic chromosome surface (PubMed:27362226). Associates with satellite DNA in G1 phase (PubMed:9510506). Binds tightly to chromatin in interphase, chromatin-binding decreases in mitosis when it associates with the surface of the condensed chromosomes (PubMed:15896774, PubMed:22002106). Predominantly localized in the G1 phase in the perinucleolar region, in the later phases it is also detected throughout the nuclear interior, being predominantly localized in the nuclear matrix (PubMed:22002106).
  • Target information above from: UniProt accession P46013 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4288 Human
    • Omim: 176741 Human
    • SwissProt: P46013 Human
    • Unigene: 689823 Human
    • Unigene: 80976 Human
    • Alternative names

      • Antigen identified by monoclonal antibody Ki 67 antibody
      • Antigen identified by monoclonal antibody Ki-67 antibody
      • Antigen KI-67 antibody
      • Antigen KI67 antibody
      • Antigen Ki67 antibody
      • KI67_HUMAN antibody
      • KIA antibody
      • Marker of proliferation Ki-67 antibody
      • MIB 1 antibody
      • MIB antibody
      • MKI67 antibody
      • PPP1R105 antibody
      • Proliferation marker protein Ki-67 antibody
      • Proliferation related Ki 67 antigen antibody
      • Protein phosphatase 1 regulatory subunit 105 antibody
      • RP11-380J17.2 antibody
      see all

    Images

    • Immunocytochemistry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
      Immunocytochemistry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)

      ab197234 staining Ki67 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab197234 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).

    • Immunocytochemistry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
      Immunocytochemistry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)

      ab197234 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab197234 at 1/100 dilution (shown in green) and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Flow Cytometry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
      Flow Cytometry - Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
      Overlay histogram showing HeLa cells stained with ab197234 (red line). The cells were fixed with 80% methanol (5 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 (ab197234, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG [EPR25A] Alexa Fluor® 488 (ab199091) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)
      Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] (ab197234)

    Protocols

    • Flow cytometry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (12)

    Publishing research using ab197234? Please let us know so that we can cite the reference in this datasheet.

    ab197234 has been referenced in 12 publications.

    • Li Y  et al. Low-abundance mutations in colorectal cancer patients and healthy adults. Aging (Albany NY) 12:808-824 (2020). PubMed: 31927530
    • Wang L  et al. Role of lncRNAHCP5/microRNA-525-5p/PRC1 crosstalk in the malignant behaviors of ovarian cancer cells. Exp Cell Res N/A:112129 (2020). PubMed: 32511950
    • Huang C  et al. ERK1/2-Nanog signaling pathway enhances CD44(+) cancer stem-like cell phenotypes and epithelial-to-mesenchymal transition in head and neck squamous cell carcinomas. Cell Death Dis 11:266 (2020). PubMed: 32327629
    • Melnik D  et al. Dexamethasone Inhibits Spheroid Formation of Thyroid Cancer Cells Exposed to Simulated Microgravity. Cells 9:N/A (2020). PubMed: 32033410
    • Zeng A  et al. Betulinic acid induces apoptosis and inhibits metastasis of human colorectal cancer cells in vitro and in vivo. Bioorg Med Chem 27:2546-2552 (2019). PubMed: 30910472
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Immunocytochemistry abreview for Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 488)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cell (Melanoma cells)
    Specification
    Melanoma cells
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 09 2020

    Immunohistochemistry (Frozen sections) abreview for Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 488)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Colon)
    Permeabilization
    No
    Specification
    Colon
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 09 2020

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