Lumit® hKi-67 Immunoassay for Cell Proliferation

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No-Wash Assay for Ki-67 Detection​

  • Increase confidence in your data by tracking a well-known marker of cell proliferation​
  • Decrease prep time using a fast, no-wash, add-and-read protocol​
  • Make quicker decisions by using an earlier readout of proliferation​
  • Available through our Early Access program

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Catalog number selected: CS3076A01

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Lumit® hKi-67 Immunoassay for Cell Proliferation
100 assays
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Assess Cell Proliferation in Less Than Two Hours with “Add-Mix-Measure” Assay

Cell proliferation is a fundamental parameter of cell-based research. Investigators currently monitor cell proliferation with methods that can be affected by changes in metabolism, require cumbersome wash steps or use indirect markers of proliferation, radioactive material and specialized equipment.

The Lumit® hKi-67 Immunoassay for Cell Proliferation is an easy add-mix-measure plate-based assay for tracking Ki-67, a well-known marker of cell proliferation, without washing steps or specialized detection equipment. A reader capable of detecting luminescence is the only instrument required.

With this assay, researchers can generate more data with higher confidence and reduce prep work and time to results.

How the Lumit® hKi-67 Immunoassay Works

Primary antibodies to Ki-67 were selected for their specific and sensitive detection and labeled with the LgBiT and SmBiT subunits of NanoBiT® Luciferase. In the presence of Ki-67, the subunits are brought together to form an active luciferase enzyme. Addition of optimized substrate generates a bright luminescent signal proportional to Ki-67 levels.

Illustration showing how the Lumit® hKi-67 Immunoassay works.

Avoid Tedious Wash Steps with Simple Protocol​

Illustration of the Lumit® hKi-67 Immunoassay protocol.

Broad Linear Range and Specificity of Lumit® hKi-67 Detection

The broad linear detection range of the Lumit® hKi-67 Immunoassay covers expected Ki-67 levels for most cell models and is specific to Ki-67 detection.

Line graph showing the specificity of the Lumit® hKi-67 Immunoassay detection.
Line graph showing the broad linear range of the Lumit® hKi-67 Immunoassay.

Measure Decreases in Cell Proliferation

Treatment of HCT 116 cells with nutlin-3A did not induce cytotoxicity, as determined using fluorescent CellTox™ Green Assay.
24 hours after treatment with nutlin-3A, Ki-67 expression is reduced in HCT 116, along with reduced viable cell number, which was assessed using the Lumit® hKi-67 Immunoassay and CyQuant™ Direct assay, respectively.
48 hours after treatment with nutlin-3A, Ki-67 expression is reduced in HCT 116, along with reduced viable cell number, which was assessed using the Lumit® hKi-67 Immunoassay and CyQuant™ Direct assay, respectively.

Treatment of HCT 116 cells (10,000/well, 96-well plate) with increasing concentrations of the antiproliferative agent nutlin-3A did not induce cytotoxicity and reduced Ki-67 expression levels in a dose-dependent manner. Ki-67 levels were assessed with the Lumit® hKi-67 Assay, while changes in viable cell number were determined with CyQUANT™ Direct in a parallel assay plate.


Measure Increases in Cell Proliferation

Treatment of CD8+ T cells with IL-2; a CD3/CD28 T Cell activator reveals only modest T cell proliferation.
Treatment of CD8+ T cells with IL-2; a CD3/CD28 T Cell activator reveals significant upregulation of Ki-67, as measured using the Lumit® hKi-67 Immunoassay.

Human CD8+ T cells (StemCell Tech) plated at 80,000 cells per well were treated with CD3/CD28 T cell activator with (orange) and without (blue) 10ng/ml IL-2 for 48 hours. Upregulation of Ki-67 is observed with the Lumit® hKi-67 Immunoassay before T cell proliferation (which begins >72 hours after activation), demonstrating use of this assay as an early indicator of proliferation.

Need to differentiate between antiproliferative effects and cell death?​

Quickly and easily assess cell proliferation with the Lumit® hKi-67 Immunoassay for Cell Proliferation, a homogeneous cell-based assay for the detection of hKi-67, a key marker of cell proliferation.

Download the Scientific Poster
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Blog Article: Rethinking Cell Proliferation Assays

The innovative Lumit® hKi-67 Immunoassay for Cell Proliferation tracks a marker that is independent of the metabolic state of cells, enabling researchers to be more confident in their assessment of proliferation.

Read About This Study

Specifications

What's in the box?

Item Part # Size

Lumit® Detection Substrate C

CS3076A02 1 × 480μl

Lumit® Immunoassay Buffer C (10X)

CS3076A03 1 × 1.8ml

Lumit® Lysis Buffer II (10X)

CS3076A04 1 × 1.3ml

Human Ki-67 Protein, Partial

CS3076A05 1 × 25μl

Anti-hKi-67 mAb-LgBiT, 400X

CS3076A06 1 × 60μl

Anti-hKi-67 mAb-SmBiT, 400X

CS3076A07 1 × 60μl

CellTox™ Green Dye, 1,000X

G873A 1 × 20μl

Certificate of Analysis

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Storage Conditions

AA

Patents and Disclaimers

U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.

U.S. Pat. Nos. 9,797,889, 9,797,890, 10,107,800 and 10,648,971 and other patents and patents pending.

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