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細胞增殖(cell proliferation)&細胞毒性(cytotoxicity assays)測定-Cell Counting Kit-8 (CCK-8)

Cell Counting Kit-8 (CCK-8)為高靈敏度檢測細胞存活率的檢測套組。 單一劑型操作容易、水溶性更佳、具高靈敏度及毒性極小，檢測時間只需1小時 。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黃色甲瓚產物（Formazan dye）。Cell Counting Kit-8 (CCK-8) 是非放射性的，對細胞的毒性極小，完全可以測完細胞生長或藥物毒性後，再繼續用細胞進行下游的實驗。Cell Counting Kit-8 (CCK-8) 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。貨號BMU106-EN 

CCK-8檢測原理: 
CCK-8試劑中含有WST-8，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。可應用於細胞增殖(cell proliferation)、細胞毒性(cytotoxicity assays)測定。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。

CCK-8產品特點:
CCK-8為高靈敏度檢測細胞存活率的檢測套組。單一劑型(Ready-to-use CCK-8 solution)操作容易、反應物水溶性更佳、 毒性極小，且檢測時間只需1小時。 靈敏度高。傳統細胞存活試驗有MTT試驗，MTT的全名是3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide；是一種黃色、接受氫離子的化合物。需要DMSO將甲䐶溶解出來之後，再利用吸光度的測定以評估有多少細胞存活，MTT需花費4小時才能得到的結果，步驟較為繁瑣。而Cell Counting (CCK-8) kit只需1小時，還可以應用於細胞生長和毒性分析。Cell Counting (CCK-8) kit 對細胞的毒性極小，完全可以測完細胞生長或藥物毒性後再繼續用細胞進行下游實驗。

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). 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。

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.

 

產品貨號	產品品項	反應	測量儀器
貨號KTA1001	活細胞和死細胞雙染色試劑盒	100個反應	活細胞綠色螢光染劑Ex/Em = 488/530 nm; 死細胞紅色螢光染料(Ex/Em = 535/617)
貨號 KTA2010	TUNEL細胞凋亡檢測試劑盒	50個反應	螢光酶標儀、螢光顯微鏡或流式細胞儀。FITC通道（Ex / Em = 490nm / 520 nm）
貨號 KTC011001	CCK-8細胞增殖和細胞毒性試劑盒	500個反應	吸收光450nm
貨號KTC4003	TraKine™粒線體染色	50個反應/250個反應	Ex / Em = 490/523 nm增殖(proliferating)和非增殖細胞(non-proliferating cells)
貨號KTC4100	Pro活細胞微管蛋白染色	50個反應/250個反應	Ex/Em = 500/520 nm
貨號KTC4001	Abbkine 細胞膜染色	100個反應/500個反應	Ex/Em = 484/501 nm
貨號BMD00063	DAPI Staining Solution	10MG/100MG	Ex/Em: 358/461 nm (with DNA)
貨號KTC4210	活細胞Lysosome染色	50個反應/250個反應	deep red fluorescent dye , Ex/Em:650/665nm
貨號KTA3030	細胞衰老檢測試劑盒（β-半乳糖苷酶）	100個反應	光學顯微鏡
貨號KTA1020	CCK-8細胞增殖和細胞毒性試劑盒	1000個反應/10,000個反應	吸收光450nm
貨號KTA2030	細胞增殖EdU Image試劑盒 (BrdU替代法)	100個反應/500個反應	AbFluor 488 azide (Ex/Em = 501/525 nm)
貨號KTB1400	一氧化Dan（NO）檢測試劑盒	96個反應/ 192個反應	colorimetric (540 nm) 檢測範圍：1-100 µM
貨號KTB1070	CheKine™黃嘌呤氧化酶檢測	96個反應/ 480個反應	OD 450 nm 線性範圍15.6 -500 mU/
貨號KTD3001	BCA蛋白定量試劑盒	2000個反應/ 5000個反應	OD 562 nm 相容性 5 % SDS, 5 % Triton X-100, 5 % Tween-20, 60, 80 50 to 1000 µg/ml.
貨號KTB1110	乳酸脫氫酶（LDH）測定試劑盒	96個反應/ 480個反應	λmax= 450 nm 1- 20 U/mL of LDH
貨號PRP100153	IGF-I重組蛋白	200UG/1MG	human IGF1 isoform 1 (P05019-1) (Gly 49-Ala 118) > 95 % as determined by SDS-PAGE
貨號 KTA2020	細胞週期染色檢測	100個反應/500個反應	Ex/Em=535/615 nm.
貨號BMC21400	PurKine™內毒素去除預裝離心柱	1ml×5 3mlx5	More than 2,000,000 EU/mL to eliminate >99% of endotoxins.
貨號KTA1200	CheKine™蛋白羰基比色測定試劑盒	48個反應/ 96個反應	吸收光370 nm
貨號KTB1080	超氧陰離子清除測定試劑盒	48個反應/ 96個反應	Superoxide anion scavenging rate D% =（∆∆ODControl – ∆∆ODSample）÷ ∆∆ODControl ×100%
貨號KTB1090	清除氫氧自由基能力測定試劑	48個反應/ 96個反應	吸收光 536nm Hydroxyl radical scavenging rate D% =（ASample – AControl）÷（ABlank – AControl）×100%
貨號KTB2220	TC總膽固醇檢測試劑盒	48個反應/ 96個反應	500nm Standard: 5 μmol/mL
貨號KTB1510	CheKine™尿酸（UA）比色分析試劑盒	48個反應/ 96個反應	吸收光505 nm Standard: 5 μmol/mL.
貨號KTB1300	Glucose葡萄糖測定試劑盒	96個反應/ 192個反應	吸光值630nm 4- 300 mg/dL)
貨號KTB1100	Lactate  乳酸檢測試劑盒	96個反應/ 480個反應	colorimetric (450 nm) 0.03 mM-2 mM
貨號KTB1700	組織和血液鹼性磷酸酶（ Alkaline Phosphatase, AKP / ALP）比色測定	96個反應/ 96 X5盤	吸收光510 nm
貨號KTB1500	總抗氧化劑能力TAC分析	96個反應/ 480個反應	593 nm 偵測範圍: 0.15-3 mM
貨號KTA0002	Annexin V-AbFluor™488細胞凋亡檢測試劑盒	20個反應/50個反應/100個反應	Annexin V- AbFluor™ 488: Abs/Em = 491/517 nm; PI: Abs/Em = 535/617 nm (with DNA)
貨號KET7013	EliKine™ Rat IL-6 ELISA Kit試劑盒	96個反應/ 96 X5盤	450 nm 偵測範圍: 250pg/mL-16000 pg/mL, 最低靈敏度100 pg/mL
貨號KET0003	雌二醇（Estradiol , E2）試劑盒	96個反應/ 96 X5盤	450 nm 偵測範圍40 pg/ml-1500 pg/ml
貨號KET6032	TNF–α Alpha型腫瘤壞死因子 ELISA試劑盒	96個反應/ 96 X5盤	450 nm 7.8 pg/ml-500 pg/ml
貨號KET6033	VEGF血管內皮生長因子ELISA試劑盒	96個反應/ 96 X5盤	450 nm 偵測範圍: 31.25 pg/mL-2000 pg/mL
貨號KET7005	白介素-1β（Interleukin-1β, IL-1β）ELISA試劑盒	96個反應/ 96 X5盤	Microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm. 偵測範圍: 15.6 pg/ml-1000 pg/ml
貨號KTE70003	小鼠黃嘌呤脫氫酶/氧化酶（XDH）ELISA試劑盒	96個反應/ 96 X5盤	450 nm 偵測範圍: 22.5 ng/L-360 ng/L, 最低靈敏度1.0 ng/L。
貨號 A02051HRP	His-tag標籤多株抗體-HRP	50ul/200ul	WB (1:5000).
貨號A02170MGB	Anti-V5 Tag Mouse Monoclonal Antibody (11D5), 磁珠	100ul/400ul/2ml	GKPIPNPLLGLDST epitope
貨號KTA4001	JC-1線粒體膜電位測定試劑盒	100個反應/ 500個反應	（聚結的JC-1，Ex / Em = 585/590 nm）發出綠色螢光（JC-1單體，Ex / Em = 510/527 nm） JC-1 Stain CCCP (10 mM)  Assay Buffer (5×)
貨號KTB1600	穀胱甘肽(Glutathione)檢測	96個反應	412 nm  2-200 µg/mL
貨號BMD00063	DAPI Staining Solution	1MG/10MG/100MG	λEx/λEm: 358/461 nm (with DNA)
貨號KTP3001	細胞核/細胞質分離試劑盒	50個反應/ 200個反應	Cytoplasmic Solution A (CES A) Cytoplasmic Solution B (CES B) • Nuclear Extraction Solution (NES) DTT (500X) Protease Inhibitor (100X)
貨號BMD00064-50UG	Calcein AM鈣黃綠素活細胞螢光染劑	50ug/1mg/10mg	λEX/λEm: 494/517 nm (pH 8) (calcein)
貨號 KTB1660	硫氧還蛋白過氧化物酶（TPX）檢測	96個反應	240nm
貨號KTB1020	NAD/NADH 分析試劑盒	96個反應/ 480個反應	OD565 nm 96 孔板測定中的檢測限值為 0.78 μM 線性最高 50 μM NAD+/NADH。
貨號KTB1070	CheKine™黃嘌呤氧化酶檢測	96個反應/ 480個反應	OD 450 nm 線性範圍15.6 -500 mU/mL
貨號KTB1040	Catalase過氧化氫酶活性檢測試劑盒	96個反應/ 480個反應	OD 540 nm 2-75 µM
貨號KTB1060	過氧化物酶（Peroxidases）分析試劑盒	96個反應	OD 664 nm 線性範圍為15.6-1000 µU/mL
貨號KTB1050	脂質過氧化檢測試劑盒(MDA)	96個反應	OD 532 nm 1-100 µM/L
貨號KTB1030	超氧化物歧化酶（SOD）活性測定試劑盒	96個反應	200-3.13 U/ml
貨號KTA1030	LDH細胞毒性測定試劑盒	96個反應/ 480個反應	OD 565 nm 10,00-100,0000 cells per well
貨號KTB3010	CheKine™ 花青素還原酶（ANR）活性比色測定試劑盒	48個反應/ 96個反應	absorption at 340 nm
貨號KTB1542	Tannase(TAN) 活性比色測定試劑盒	48個反應/ 96個反應	absorbance at 270 nm 0.3125-20 μmoL/mL
貨號KTB1541	CheKine™ 單寧比色測定試劑盒 | CheKine™ Tannin Colorimetric Assay Kit	48個反應/ 96個反應	absorbance at 760 nm 0.0156-1 mg/mL
貨號KTB2250	CheKine™ 微量高密度脂蛋白膽固醇 (HDL-C) 檢測試劑盒	48個反應/ 96個反應	Absorption peak at 500nm Detection range: 0.078-5 mmol/L(The Detection range corresponds to the standard, and the actual content of sample is 0.156-10 mmol/L）
貨號KTB2260	CheKine™ 微量低密度脂蛋白膽固醇 (LDL-C) 檢測試劑盒	48個反應/ 96個反應	Absorption peak at 500nm Detection range: 0.078-5 mmol/L(The Detection range corresponds to the standard, and the actual content of sample is 0.117-7.5 mmol/L）
貨號KTB1290	CheKine™ 烏頭酸酶 (ACO) 活性比色測定試劑盒	48個反應/ 96個反應	absorption at 340 nm.
貨號KTB1560	醇醯基轉移酶（AAT）活性比色檢測試劑盒	48個反應/ 96個反應	absorption at 412 nm.
貨號KTB1390	CheKine™ 澱粉分支酶 (SBE) 活性比色測定試劑盒	48個反應/ 96個反應	absorption at 660 nm
貨號PRP1012	Human IL-6 protein重組蛋白	5ug/20ug/100ug/1mg	human IL-6 (NP_000591.1) (Val 30-Met 212)/ E. coli
貨號PRP100273	Human IL-4 protein重組蛋白	20ug/100ug/1mg	ED50 for this effect is 0.05-0.2 ng/mL. The specific activity of Recombinant Human IL-4 is approximately 3.2 × 10E4 IU/μg.
貨號PRP100159	Human EGF protein重組蛋白	100ug/1mg
貨號KTB1041	CheKine™ 過氧化氫 (H2O2) 檢測試劑盒	96個反應/ 480個反應	OD 580 nm : 1 µM-100 µM
貨號KTB1210	CheKine™ 超氧陰離子比色測定試劑盒	48個反應/ 96個反應	absorption peak at 540nm 0.0125-0.5 mg/mL