細胞增殖、活性及毒性分析系列商品 Cell Proliferation, Viability & Cytotoxicity

細胞增殖速率(Cell proliferation rates)或存活力水平(viability levels)；是細胞健康的良好指標；增殖(Proliferation)或存活力分析(viability analysis)，對於細胞生長和分化研究至關重要，並且通常與代謝分析結合；代謝活性(Metabolic activity)通常用作生存力指標。增殖(Proliferation)為測量細胞因子(cytokines)、生長因子(growth factors)於細胞動力學簡易之方法。另一方面，評估化合物之細胞毒性也是藥物開發中的關鍵步驟；評價化合物毒性和藥物開發期間，對腫瘤細胞生長的抑制。特定實驗中的細胞活力可以取決於從細胞群的氧化還原電位、ATP / ADP水平、細胞膜的完整性，到細胞酶活性，例如LDH和esterases酯酶的活性之不同標準。因此，可以定量的生物標誌物包括ATP，NADH，半胱天冬酶，LDH，和活細胞和死細胞蛋白酶。

細胞活力/細胞毒性測定可以分為4種類型：

1）功能性測定，例如ATP水平。

2）細胞學/膜滲漏測定，例如LDH測定。

3）線粒體測定，例如使用Resazurin的測定。

4）基因組/蛋白質組測定

 CCK-8細胞增殖和細胞毒性試劑盒( 高靈敏度)  | SuperKine™ Maximum Sensitivity Cell Counting Kit-8 (CCK-8)貨號BMU106-EN 

Wst-8 was an upgrade of MTT and was reduced by mitochondrial dehydrogenase in the presence of electron-coupled reagents to form a highly water-soluble orange dirty product (Formazan).

Cell Counting Kit-8 (CCK-8) 通過使用高度水溶性四唑鹽-WST-8.[2-(2- methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5- 允許非常方便的檢測(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt] 在電子介體存在下還原時產生水溶性甲染料。Cell Counting Kit-8 (CCK-8) 是非放射性的，允許進行靈敏的比色測定，以確定細胞增殖(cell proliferation)和細胞毒性測定(cytotoxicity assays)中的活細胞數量。 Cell Counting Kit-8 (CCK-8) 使用試劑 WST-8 作為底物。高度水溶性的四唑鹽 WST-8 可被細胞脫氫酶裂解為可溶性甲臢。這種生物還原是透過代謝活躍細胞中脫氫酶產生的 NADPH 或 NADH 來完成的。甲臢染料產品的量與培養物中代謝活躍細胞的數量直接相關。WST-8 在細胞中被脫氫酶還原，生成橙色產物（甲臢），可溶於組織培養基。細胞中脫氫酶產生的甲臢染料的量與活細胞的數量成正比。 WST-8 生產的產品 (formazan) 是水溶性的，不需要有機溶劑(organic solvents )或同位素(isotopes)。formazan穩定安全。使用 CCK-8 的檢測靈敏度高於使用其他四唑鹽（如 MTT、XTT、MTS 或 WST-1）的檢測。可應用於細胞增殖(cell proliferation)、細胞毒性(cytotoxicity assays)測定。

Publications(23)

使用此試劑盒所獲得的結果顯示出比使用MTT、MTS、XTT或WST-1等四唑化合物的方法更高的靈敏度，且時間更短且毒性更小。The detection sensitivity using CCK-8 is higher than assays using other tetrazolium salts such as MTT, XTT, MTS or WST-1.

圖1 SuperKine™ Maximum Sensitivity Cell Counting Kit-8 (CCK-8) (Cat # : BMU106-EN) 與國際三大品牌A、B、C檢測不同活細胞數HEK293細胞的比較.結果表明，SuperKine™ Maximum Sensitivity Cell Counting Kit-8 (CCK-8) 具有最佳的靈敏度。

使用 CCK-8 獲得的細胞增殖或細胞毒性測定結果顯示出比使用四唑化合物（如 MTT、MTS 或 WST-1）的方法更高的靈敏度。

只有使用 CCK-8 (WST-8)，才能在不殺死細胞的情況下進行連續細胞培養。以下是觀察每種試劑孵育後細胞毒性的圖示。

Cell Number Determination Protocol

1.Inoculate cell suspension (100μL/well) in a 96-well plate.Pre-incubate the platein ahumidified in cubator (at 37℃, 5%CO2).

2.Add 10μL of the CCK-8 solution to each well of the plate.Becareful not to introduce bubbles to the wells, since they interfere with the OD reading.

3.Incubate the plate for 1-4h in the incubator.The incubation time depends on thee xperimental conditions such as cell type and cell density;

4.Measure the absorbance at 450nm using amicroplatereader.

Cell Proliferation and Cytotoxicity Assay

Protocol

1.Dispense 100μL of cell suspension(5000 cells/well) in a 96-well plate. Pre-incubate the plate for 24 h in ahumidify ingcarbon dioxideincubator(at37℃,5%CO2).

2.Add 1-10μL of various concentrations of substances to betested to the plate.

3.Incubate the plate for anappropriatelengthoftime(e.g.,6,12,24or48h) in the incubator.

4.Add 10μL of CCK-8 solution to each well of the plate. Becareful not to introduce bubbles to th ewells, since they interfere with the OD reading.

5.Incubate the plate for 1-4h in the incubator.The incubation timed ependson thee xperimental conditions such ascelltypeand celldensity;

6.Measure the absorbance at 450nm using a microplatereader.

活細胞和死細胞雙染色套組  |  CCK-8細胞增殖和細胞毒性試劑盒  | LDH細胞毒性測定試劑盒 | 細胞衰老檢測試劑盒（β-半乳糖苷酶 | 細胞增殖EdU Image試劑盒 

Product name	SuperKine™ Maximum Sensitivity Cell Counting Kit-8 (CCK-8)
Applications notes	Wst-8 was an upgrade of MTT and was reduced by mitochondrial dehydrogenase in the presence of electron-coupled reagents to form a highly water-soluble orange dirty product (Formazan).
Formulation	Liquid solution
Kit components	Ready-to-use CCK-8 solution
Features & Benefits	•High sensitivity: More accurate detection results; •Good stability: Can be stored at -20℃ for more than 2 years; •High safety: Less cytotoxicity, almost no impact on subsequent experiments.
Storage instructions	Store at 4℃ for 12 months away from light, at least 24 months at -20℃
Shipping	Blue ice
Precautions	The reagent is only used in the field of scientific research, not suitable for clinical diagnosis or other purposes.

LDH 細胞毒性檢測試劑盒 | LDH Cytotoxicity Assay Kit貨號KTA1030

LDH Cytotoxicity Assay Kit

Detect lysis of 1000-100,0000 cells per well.

LDH (Lactate dehydrogenase, LDH)測定，也稱為 LDH 釋放測定，是一種細胞死亡(cell death)/細胞毒性測定(cytotoxicity assay)，用於評估細胞群中質膜損傷(plasma membrane damage)的水平。乳酸脫氫酶 (Lactate dehydrogenase, LDH)是一種穩定的酶，存在於所有細胞類型中，在質膜((plasma membrane )受損後迅速釋放到細胞培養基中。 LDH 是用於運行細胞毒性(cytotoxicity assay)測定的最廣泛使用的標記物。

細胞死亡(Cell death)或細胞毒性(cytotoxicity)通常通過質膜損傷(plasma membrane damage)的量化來評估。 LDH 是一種穩定的酶，存在於所有細胞類型中，並在質膜(plasma membrane)受損後迅速釋放到細胞培養基中。 因此，LDH 是細胞毒性(cytotoxicity)研究中使用最廣泛的標誌物。細胞毒性檢測試劑盒(Abbkine LDH Cytotoxicity Assay Kit, KTB1030)為細胞毒性(cytotoxicity)研究提供了一種簡單易行的比色檢測方法(colorimetric assay)。 該測定基於在 NADH 偶聯酶促反應(NADH-coupled enzymatic reaction)中將四唑鹽 MTT (tetrazolium salt MTT) 還原為 MTT 的還原形式，其在 565 nm 處具有最大吸收。 生成顏色的強度與裂解的細胞數量直接相關。 在典型的細胞毒性試驗(typical cytotoxicity assay)中，靶細胞(target cells)與細胞毒性化學試劑(cytotoxic chemical agent)或細胞毒性細胞（NK 細胞、細胞毒性 T 細胞）一起培養，以誘導靶細胞 (target cells)死亡和 LDH 釋放。 將含有 LDH 的上清液轉移到新的 96 孔測定板的孔中，並與 LDH 反應溶液混合。 在室溫下孵育 30 分鐘後，使用讀板器讀取 565nm (A565) 處的吸光度。

Abbkine LDH Cytotoxicity Assay Kit provides a simple and easy colorimetric assay for the study of cytotoxicity. This assay is based on the reduction of the tetrazolium salt MTT in a NADH-coupled enzymatic reaction to a reduced form of MTT which exhibits an absorption maximum at 565 nm. The intensity of the generated color correlates directly with the cell number lysed. In a typical cytotoxicity assay, target cells are cultured with a cytotoxic chemical agent or a cytotoxic cell (NK cell, cytotoxic T cells) to induce target cell death and LDH release. The LDH-containing supernatants are transferred to wells of a new 96-well assay plate and mixed with the LDH Reaction solution. After an incubation of 30 minutes at room temperature, the absorbance at 565nm (A565) is read using a plate reader. Abbkine LDH

Kit components	• Assay Buffer • Lactic Acid Solution • MTT Solution • PES Solution • LDH Positive Control • NAD+ Solution • Triton X-100 (10%)
Features & Benefits	• Measures cell death in response to chemical compounds or environmental factors. • LDH release from the cytosol into the medium catalyzes a color-forming reaction. • Detect lysis of 10,00-100,0000 cells per well.
Usage notes	Performing an initial titration experiment to determine the optimal number of cells per well of the target cell you plan to use.
Storage instructions	Store at -20°C and Keep in dark. The Kit has a storage time of 6 months from receipt.
Shipping	Gel pack with blue ice.
Precautions	The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.

LDH Cytotoxicity Assay Kit (CAT#KTA1030) 是用於細胞毒性檢測的。CheKine™ Micro Lactate Dehydrogenase (LDH) Assay Kit  (CAT#KTB1110)主要是用於細胞或者組織內LDH酶活定量的。