Cell Counting Kit-8
细胞增殖与毒性检测试剂盒
货号:AKCE001-2
规格: 500T
检测设备:-
可检测样本数:500 Samples
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Product Information
Cell Counting Kit-8
-20℃ Dry Ice Transportation
  • 检测样本量:500 Samples
  • 主要检测设备及配套:酶标仪/96孔板
  • 预计测定时间:-
  • 试剂储存条件:按照标签要求储存
  • 需自备试剂:
Detection Principle

Cell Counting Kit-8 (CCK-8) 作为MTT法的替代方法,可对细胞增殖和细胞毒性进行快速、高灵敏度、无放射性的比色检测,广泛应用于抗肿瘤药物筛选、生物活性因子检测、细胞增殖或毒性测定、肿瘤药敏等试验。

CCK-8试剂无需预配即可直接加入细胞样品中,试剂中含有WST-8 [2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium,monosodium salt],WST-8在电子耦合试剂(1-Methoxy PMS)的作用下被线粒体内的脱氢酶还原生成高度水溶性的橙黄色Formazan,产物数量与活细胞数量呈线性关系,通过测定450 nm处吸光值的变化即可反映活细胞的数量。

  • 检测方法: CCK-8
  • 检测波长: 450 nm
  • 信号响应: 递增型
Notices

①CCK-8的培养时间一般为1-4小时,需要摸索接种细胞的数量和加入CCK-8试剂后的培养时间,CCK-8的最佳反应时间以具体显色最佳时间为准;

②孵育时间因细胞类型和数量而异;白细胞着色较弱,可能需要较长的孵育时间或大量细胞;

③当使用标准96孔板时,贴壁细胞的最小接种量为1000个细胞/孔(100 μL培养基);检测白细胞时的灵敏度相对较低,推荐接种量应不低于2500个细胞/孔(100 μL培养基);如果要使用24孔板或6孔板,请先计算每孔相应的接种量,并按照每孔培养基总体积的10%加入CCK-8溶液;

④如果没有450 nm滤光片,可以使用430-490 nm之间的滤光片,但450 nm检测灵敏度最高。如果您需要进行双波长测定,650 nm处的吸光度可作为参考波长进行测定;

⑤培养基中的酚红不会影响实验结果,酚红的吸光度可以通过扣除空白孔中本底的吸光度而消去;

⑥本产品可以检测E.coli,但不能检测酵母细胞;

⑦如果您需要对CCK-8溶液进行无菌处理,请使用0.2 μm滤膜过滤除菌;

⑧使用96孔板进行检测时,96孔板外圈孔位易蒸发,如果细胞培养时间较长,可以选择弃用外圈孔位,或改加等量的PBS、水或培养液作为填充液。

注: 为保证结果准确且避免试剂损失,测定前请仔细阅读说明书(以实际收到说明书为准),确认试剂储存和准备是否充分,操作步骤是否清楚,且务必取2-3个预期差异交的样本进行预测定,过程中问题请您及时与工作人员联系。
Product Citation

[1] Du J, Yu L, Yang X, et al. Regulation of NCOA4-mediated iron recycling ameliorates paraquat-induced lung injury by inhibiting ferroptosis[J]. Cell Communication and Signaling, 2024, 22(1): 146. (IF 8.4)

[2] Li H, Li Y, Yu Y, et al. GSH exhaustion via inhibition of xCT-GSH-GPX4 pathway synergistically enhanced DSF/Cu-induced cuproptosis in myelodysplastic syndromes[J]. Free Radical Biology and Medicine, 2024. (IF 7.1)

[3] Du P, Song J, Qiu H, et al. Polyphenols Extracted from Shanxi-Aged Vinegar Inhibit Inflammation in LPS-Induced RAW264. 7 Macrophages and ICR Mice via the Suppression of MAPK/NF-κB Pathway Activation[J]. Molecules, 2021, 26(9): 2745.(IF 4.412)

[4] Song J, Qiu H, Du P, et al. Polyphenols extracted from Shanxi‐aged vinegar exert hypolipidemic effects on OA‐induced HepG2 cells via the PPARα‐LXRα‐ABCA1 pathway[J]. Journal of food biochemistry, 2022, 46(2): e14029.(IF 3.654)

[5] Song J, Qiu H, Geng X, et al. Immunomodulatory effects of vinegar‐egg juice: Potential pharmacological effects of a traditional Chinese food remedy?[J]. Journal of Food Biochemistry, 2022, 46(12): e14418.(IF 3.654)

[6] Yu B, Li Q, Sun X, et al. O‐GlcNAcylation of TLR4 inhibits osteogenic differentiation of periodontal ligament stem cells[J]. Journal of Periodontal Research, 2023.(IF 3.5)

[7] Lou Y, Huang K, Xu B, et al. METTL14 plays an oncogenic role in NSCLC by modulating ferroptosis and the m6A modification of GPX4[J]. Archives of Physiology and Biochemistry, 2024: 1-12. (IF 2.5)

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