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AB215226

Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610]

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(2 Publications)

Rabbit Recombinant Monoclonal Ki67 antibody - conjugated to Alexa Fluor® 555. Suitable for ICC, Flow Cyt (Intra) and reacts with Human samples. Cited in 2 publications.

View Alternative Names

Proliferation marker protein Ki-67, Antigen identified by monoclonal antibody Ki-67, Antigen KI-67, Antigen Ki67, MKI67

3 Images
Immunocytochemistry - Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610] (AB215226)
  • ICC

Lab

Immunocytochemistry - Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610] (AB215226)

ab215226 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 ab215226 at 1/100 dilution (pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 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® 555 Anti-Ki67 antibody [EPR3610] (AB215226)
  • ICC

Lab

Immunocytochemistry - Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610] (AB215226)

ab215226 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 overnight at +4°C with ab215225 at a 1/500 dilution (pseudocolored in green) and ab195884, Rat monoclonal [YOL1/34] to Tubulin (Alexa Fluor® 647), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 4% formaldehyde (10 min).

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

Flow Cytometry (Intracellular) - Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610] (AB215226)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 555 Anti-Ki67 antibody [EPR3610] (AB215226)

Overlay histogram showing HeLa cells stained with ab215226 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab215226, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor® 555 (ab208569) 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 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR3610

Isotype

IgG

Conjugation

Alexa Fluor® 555

Excitation/Emission

Ex: 555nm, Em: 565nm

Carrier free

No

Reacts with

Human

Applications

ICC, Flow Cyt (Intra)

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICC" : {"fullname" : "Immunocytochemistry", "shortname":"ICC"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICC-species-checked": "testedAndGuaranteed", "ICC-species-dilution-info": "1/100 - 1/500", "ICC-species-notes": "<p>This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 - 20 min) and 100% methanol (5 min).</p><p>If fixing cells in 4% PFA, it is recommended to permeabilized cells with 0.1% Triton-X for 5 min.</p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/500", "FlowCytIntra-species-notes": "<p></p>" } } }
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Product details

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle|Store in the dark
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Ki67 is a nuclear protein often referred to as MKI67 with a molecular weight of approximately 345 kDa. This protein is strongly associated with cell proliferation. Scientists commonly use Ki67 staining techniques to identify and quantify proliferating cells in tissues. Ki67 is expressed in the nucleus during active cell cycle phases—G1 S G2 and M—but not in resting cells in G0 phase. It is detectable through methods such as Ki67 immunofluorescence and Ki67 flow cytometry which are important for analyzing cell division.
Biological function summary

The Ki67 protein plays an essential role in cellular proliferation processes. It does not form part of a stable complex but interacts transiently with other cell cycle-related proteins. Research indicates that Ki67 maintains the structure of the perichromosomal layer during mitosis impacting chromosome separation. It is particularly active in tissues with high cell turnover such as bone marrow and lymphoid organs.

Pathways

Ki67 influences several key cellular pathways that control cell proliferation and differentiation. The protein acts within the regulation of the cell cycle and the PI3K/AKT signaling pathway important for cell growth and survival. Within these pathways Ki67 interacts with proteins like cyclin-dependent kinases (CDKs) which regulate transitions between different phases of the cell cycle.

Ki67 has significant implications in oncology and can be used as a biomarker for cancer prognosis. Its expression levels help determine the aggressiveness of tumors especially in breast cancer and prostate cancer. High Ki67 levels correlate with poor prognosis as it indicates rapid cell division. In cancer Ki67 associates with p53 a protein that regulates the cell cycle and function as a tumor suppressor.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

The protein expressed by the MKI67 gene is required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly and associates with the surface of the mitotic chromosome, specifically the perichromosomal layer, covering a significant portion of the chromosome surface (PubMed : 27362226). It prevents chromosomes from collapsing into a single chromatin mass by creating a steric and electrostatic charge barrier due to its high net electrical charge, acting as a surfactant that disperses chromosomes and enables their independent motility (PubMed : 27362226). The protein binds DNA, with a preference for supercoiled and AT-rich DNA (PubMed : 10878551), and may play a role in chromatin organization, though it is unclear if it directly influences chromatin organization or if this is an indirect result of its function in maintaining dispersed mitotic chromosomes (Probable, PubMed : 24867636). This supplementary information is collated from multiple sources and compiled automatically.
See full target information MKI67
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Publications (2)

Recent publications for all applications. Explore the full list and refine your search

JCI insight 10: PubMed40178918

2025

Dipeptidase-1-knockout mice develop invasive tumors with features of microsatellite-unstable colorectal cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Sarah E Glass,Matthew E Bechard,Zheng Cao,Radhika Aramandla,Ping Zhao,Samuel T Ellis,Emily H Green,Elizabeth G Fisher,Ryan T Smith,Chelsie K Sievers,Maria Johnson Irudayam,Frank Revetta,M Kay Washington,Gregory D Ayers,Cody N Heiser,Alan J Simmons,Yanwen Xu,Yu Wang,Annika Windon,Martha J Shrubsole,Nicholas O Markham,Qi Liu,Ken S Lau,Robert J Coffey

Micromachines 13: PubMed35056257

2022

A Cell Culture Chip with Transparent, Micropillar-Decorated Bottom for Live Cell Imaging and Screening of Breast Cancer Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Menekse Ermis,Ezgi Antmen,Ozgur Kuren,Utkan Demirci,Vasif Hasirci
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Product promise

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