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AB253221

Human Ki67 ELISA Kit

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(4 Publications)

Human Ki67 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Ki67 with a sensitivity of 21.2 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

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

3 Images
ELISA - Human Ki67 ELISA Kit (AB253221)
  • ELISA

Supplier Data

ELISA - Human Ki67 ELISA Kit (AB253221)
Sandwich ELISA - Human Ki67 ELISA Kit (AB253221)
  • sELISA

Unknown

Sandwich ELISA - Human Ki67 ELISA Kit (AB253221)

Example of human Ki67 standard curve in 1X Cell Extraction Buffer PTR.

The Ki67 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human Ki67 ELISA Kit (AB253221)
  • sELISA

Unknown

Sandwich ELISA - Human Ki67 ELISA Kit (AB253221)

Interpolated concentrations of native Ki67 in human A431 and Hela cell extracts based on a 250 µg/mL and 62.5 µg/mL extract load respectively.

The concentrations of Ki67 were measured in duplicate and interpolated from the Ki67 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Ki67 concentration was determined to be 1,575.0 pg/mL in A431 and 1674.3 pg/mL in Hela.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 21.2 pg/mL

Range

31.25 - 2000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Primary Antibodies

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human Ki67 ELISA Kit ab253221 is a rapid single-wash 90-min Sandwich ELISA to measure Human Ki67 in cell lysate. This SimpleStep sensitivity is 21.2 pg/mL.

How the assay works

Human Ki67 SimpleStep ELISA®employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human Ki67 ELISA Kit ab253221 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-Ki67 antibody [EPR20582-139] - BSA and Azide free (Capture) ab252656
- Anti-Ki67 antibody [EPR20582-7] - BSA and Azide free (Detector) ab252656

Ki67 is a nuclear marker protein that is commonly found in active proliferating cells. Encoded by the *MKI67* gene, Ki67 is expressed in all phases of the cell cycle except for resting cells in G0 stage. Moreover, the presence of Ki67 protein is at its highest in the G2 phase and during mitosis. Studies have shown Ki67 to be an excellent antigen marker to determine the growth fraction of a given cell population. More particularly in cancer, which is subsequently correlated with the clinical course of action. The standard protein in ab253221 is an internal fragment comprising 7% of the full-length Ki67.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "1.8" }, { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4.1" } ]
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Recovery

[ { "sample": "Cell Lysate", "range": "101 - 116 %", "average": "= 108" } ]
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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab253221-1x96Tests", "assetComponentDetails": [ { "size":"1 x 6 mL", "name":"Antibody Diluent CPR2", "number":"AB253221-CMP13", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human Ki67 Capture Antibody", "number":"AB253221-CMP07", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB253221-CMP12", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB253221-CMP11", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human Ki67 Lyophilized Recombinant Protein", "number":"AB253221-CMP09", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human Ki67 Detector Antibody", "number":"AB253221-CMP08", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB253221-CMP05", "productcode":"" }, { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB253221-CMP04", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB253221-CMP10", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB253221-CMP06", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB253221-CMP03", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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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

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Target data

Protein that associates with the surface of mitotic chromosomes and acts both as a chromosome repellent during early mitosis and chromosome attractant during late mitosis (PubMed : 27362226, PubMed : 32879492, PubMed : 35513709, PubMed : 39153474). Required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly (PubMed : 27362226). During early mitosis, relocalizes from nucleoli to the chromosome surface where it forms extended brush structures that cover a substantial fraction of the chromosome surface (PubMed : 27362226). The MKI67 brush structure 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). During mitotic anaphase, the MKI67 brush structure collapses and MKI67 switches from a chromosome repellent to a chromosome attractant to promote chromosome clustering and facilitate the exclusion of large cytoplasmic particles from the future nuclear space (PubMed : 32879492, PubMed : 39153474). Mechanistically, dephosphorylation during mitotic exit and simultaneous exposure of a conserved basic patch induce the RNA-dependent formation of a liquid-like condensed phase on the chromosome surface, promoting coalescence of neighboring chromosome surfaces and clustering of chromosomes (PubMed : 39153474). Binds premature ribosomal RNAs during anaphase; promoting liquid-liquid phase separation (PubMed : 28935370, PubMed : 39153474). 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; it is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in mitotic chromosome (PubMed : 24867636).
See full target information MKI67
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Publications (4)

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

Materials today. Bio 32:101768 PubMed40290895

2025

A microfluidic electrochemical immunosensor for detection of CEA and Ki67 in 3D tumor spheroids.

Applications

Unspecified application

Species

Unspecified reactive species

Sujin Kim,Seonyeop Kim,Chanjin Ko,Wonseok Lee,Hwan Drew Kim

International journal of molecular sciences 25: PubMed38999957

2024

Effects of High-Mobility Group Box-1 on Mucosal Immunity and Epithelial Differentiation in Colitic Carcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Takamitsu Sasaki,Rina Fujiwara-Tani,Yi Luo,Ruiko Ogata,Rika Sasaki,Ayaka Ikemoto,Yukiko Nishiguchi,Chie Nakashima,Shingo Kishi,Kiyomu Fujii,Hitoshi Ohmori,Naohide Oue,Hiroki Kuniyasu

Molecules (Basel, Switzerland) 27: PubMed35889210

2022

A Novel ERK2 Degrader Z734 Induces Apoptosis of MCF-7 Cells via the HERC3/p53 Signaling Pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Shiyao Xu,Yan Xiong,Rui Yao,Rong Tian,Zhuqing Meng,Mohamed Y Zaky,Beibei Fu,Dong Guo,Lulu Wang,Feng Lin,Xiaoyuan Lin,Haibo Wu

Biomedicines 10: PubMed35453639

2022

Non-Invasive Physical Plasma Generated by a Medical Argon Plasma Device Induces the Expression of Regenerative Factors in Human Gingival Keratinocytes, Fibroblasts, and Tissue Biopsies.

Applications

Unspecified application

Species

Unspecified reactive species

Benedikt Eggers,Matthias Bernhard Stope,Jana Marciniak,Werner Götz,Alexander Mustea,James Deschner,Marjan Nokhbehsaim,Franz-Josef Kramer
View all publications
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Product promise

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