Lumit® Cell Proliferation Assay (Human Ki-67)
Ki-67 検出のためのウォッシュ不要アッセイ
- 細胞増殖の代表的マーカーを用いたデータ信頼性の向上
- 洗浄不要・迅速なアッセイプロトコルによる前処理時間の短縮
- 増殖の早期指標に基づく迅速な意思決定
- 本製品は、Early Access プログラムを通じてご利用いただけます。
Catalog Number:
Size
Catalog Number: GC1000
Catalog Number: GC1002
Catalog Number: GC1001
「添加・混合・測定」アッセイにより、2時間以内に細胞増殖を評価
Lumit® hKi-67イムノアッセイは、細胞増殖の指標としてよく知られているKi-67を追跡するための簡単な添加・混合・測定のプレートベースのアッセイで、洗浄ステップや特殊な検出装置は不要です。発光プレート－リーダー（ルミノメーター）があれば測定可能です。
このアッセイを使用することで、研究者はより高い信頼性を持つデータを生成し、準備作業と結果までの時間を短縮することができます。
Watch the video to hear how researcher Clara Gouez is using the Lumit® Cell Proliferation Assay (Human Ki-67) to advance her gastric cancer research.
Lumit® hKi-67 Immunoassay の仕組み
シンプルなワンプレートプロトコル、洗浄ステップなし！
Compared with conventional fixation-based Ki-67 flow cytometry, the assay is significantly simpler and faster, requiring no dissociation or wash steps while still producing robust and reproducible results.
Zeynep Kaya, PhD, Postdoctoral Research Fellow, Prof Andrew Beggs Group, University of Birmingham
Lumit® hKi-67検出の広いダイナミックレンジと特異性
細胞増殖の減少を測定
細胞増殖の増加を測定
Compatible with 3D Culture Methods
In 3D spheroids, decreases in hKi-67 protein levels provide a clear and early indication of antiproliferative activity with a larger response window than a metabolic measure of viable cell number. HCT116 cells (1,000 cells/well) were plated in SBio PrimeSurface 96U plates (ULA; round bottom) and grown for 3 days to form 3D spheroids. The resultant HCT116 spheroids were then treated for 24 hours with increasing concentrations of nutlin-3a. Subsequently, proliferation was assessed with the Lumit® Cell Proliferation Assay (Human Ki-67) and a metabolic activity assay in separate plates. Note: Day 4 untreated HCT116 spheroids were ~440µm in diameter.
Protocols
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
CellTox™ Green Dye, 1,000X |
G873A | 1 × 20μl |
Human Ki-67 Protein (Partial) Positive Control |
GC100A | 1 × 25μl |
Anti-hKi-67 mAb SmBiT, 400X |
GC101A | 1 × 60μl |
Anti-hKi-67 mAb LgBiT, 400X |
GC102A | 1 × 60μl |
Lumit® Immunoassay Buffer C, 10X |
VB115C | 1 × 1.8ml |
Lumit® Lysis Buffer II, 10X |
VB310C | 1 × 1.3ml |
Ki-67 Assay Substrate |
VB321A | 1 × 600μl |
Lumit® Detection Buffer B |
VB406D | 1 × 6ml |
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 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
CellTox™ Green Dye, 1,000X |
G873B | 1 × 200μl |
Human Ki-67 Protein (Partial) Positive Control |
GC100A | 1 × 25μl |
Anti-hKi-67 mAb SmBiT, 400X |
GC101A | 5 × 60μl |
Anti-hKi-67 mAb LgBiT, 400X |
GC102A | 5 × 60μl |
Lumit® Immunoassay Buffer C, 10X |
VB115C | 5 × 1.8ml |
Lumit® Lysis Buffer II, 10X |
VB310C | 5 × 1.3ml |
Ki-67 Assay Substrate |
VB321A | 5 × 600μl |
Lumit® Detection Buffer B |
VB406D | 5 × 6ml |
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 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
CellTox™ Green Dye, 1,000X |
G873B | 1 × 200μl |
Human Ki-67 Protein (Partial) Positive Control |
GC100A | 1 × 25μl |
Anti-hKi-67 mAb SmBiT, 400X |
GC101B | 1 × 600μl |
Anti-hKi-67 mAb LgBiT, 400X |
GC102B | 1 × 600μl |
Lumit® Immunoassay Buffer C, 10X |
VB115D | 1 × 18ml |
Lumit® Lysis Buffer II, 10X |
VB310D | 1 × 13ml |
Ki-67 Assay Substrate |
VB321B | 1 × 6ml |
Lumit® Detection Buffer B |
VB406E | 1 × 60ml |
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 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents and patents pending.
Resources
Featured Resource
Need to differentiate between antiproliferative effects and cell death?
Quickly and easily assess cell proliferation with the Lumit® Cell Proliferation Assay (Human Ki-67), a homogeneous cell-based assay for the detection of hKi-67, a key marker of cell proliferation.
技術記事
技術ポスター
- Poster: Homogeneous Bioluminescent Immunoassay for hKi-67: A Simple and Robust Screening Tool for Antiproliferative Agents
- Poster: Novel, Luminescent Ki-67 Immunoassay Assesses T Cell Responses
- Poster: Lumit® Immunoassays: Bioluminescent, Sensitive, and Homogeneous Analyte Detection Using Labeled Antibodies
Compare Products
|
Cell Viability (ATP) |
Cell Viability (ATP) |
3D Cell Viability (ATP) |
Real-Time Viability |
Non-Lytic Viability |
Cell Proliferation |
|
|---|---|---|---|---|---|---|
| Best Use | Routine viability & HTS screening | Established protocols using original formulation | 3D spheroids, organoids, microtissues | Kinetic monitoring over time; downstream multiplexing | Multiplex first step; cells needed for follow-up assays | True proliferation readout independent of metabolism |
| Key Decision Points | ||||||
| Measures | Viability | Viability | Viability (3D) | Viability (kinetic) | Viability | Proliferation |
| Cells alive after assay? | ✗ Lytic | ✗ Lytic | ✗ Lytic | ✓ Non-lytic | ✓ Non-lytic | ✗ Lytic |
| Multiplexing compatible? | LimitedLytic—must be terminal step | LimitedLytic—must be terminal step | LimitedLytic—must be terminal step | ✓ ExcellentNon-lytic; pair with any downstream assay | ✓ ExcellentNon-lytic; pair with Caspase-Glo®, CTG, etc. | ModerateCan multiplex with CellTox™ Green or other fluorescent readouts |
| Real-time monitoring? | ✗ Endpoint | ✗ Endpoint | ✗ Endpoint | ✓ Up to 72hRead same wells repeatedly | ✗ Endpoint | ✗ Endpoint |
| 3D culture compatible? | PartialWorks for small spheroids; use 3D version for dense structures | PartialSame as 2.0 | ✓ OptimizedEnhanced lysis for dense 3D structures | PartialSubstrate must penetrate; best for small/loose 3D models | PartialSubstrate access may be limited in dense 3D | ✓ YesDetects Ki-67 in cell lysates from any culture format |
| Assay Attributes | ||||||
| Assay Principle | ATP quantitation (luciferase/luciferin) | ATP quantitation (luciferase/luciferin) | ATP quantitation (enhanced lysis for 3D) | Metabolic reduction of pro-substrate to luciferase substrate | Live-cell protease activity (GF-AFC cleavage) | Ki-67 immunodetection via NanoBiT® complementation |
| Detection Mode | Luminescence | Luminescence | Luminescence | Luminescence | Fluorescence400Ex / 505Em | Luminescence |
| Reagent Format | Ready-to-use liquid | Buffer + lyophilized substrateRequires reconstitution | Ready-to-use liquid | 2 components(enzyme + substrate) | Single reagent | Antibody mix + detection reagent |
| Time to Result | 10min | 10min | ~30min | ContinuousFirst read: 1–2h after addition | 30min | ~2h |
| Practical Considerations | ||||||
| Plate Formats | 96, 384, 1536 | 96, 384, 1536 | 96, 384 | 96, 384 | 96, 384 | 96, 384 |
| HTS Suitability | ✓ Excellent1536-well capable; fast protocol | ✓ Excellent1536-well capable | ✓ Good | ModerateRequires kinetic reader scheduling | ✓ Good | ✓ Good |
| Sensitivity (96-well) | ~15 cells/well | ~10 cells/well | Spheroid-dependent | <100 cells/well | ~40 cells/well | Cell line-dependent |
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