操作簡(jiǎn)單:產(chǎn)品本身是液體,即開(kāi)即用;甲臜產(chǎn)物是水溶性的,不需要用有機(jī)溶劑溶解。
靈敏度高:線性范圍廣,靈敏度高,重復(fù)性好。
細(xì)胞毒性低:細(xì)胞毒性非常低,對(duì)后續(xù)實(shí)驗(yàn)沒(méi)有影響。
穩(wěn)定性好:培養(yǎng)液中酚紅和血清的存在不會(huì)對(duì)檢測(cè)造成影響。
應(yīng)用性廣:可用于大規(guī)模、高通量樣品檢測(cè)。 【詳細(xì)說(shuō)明】
CCK-8,您細(xì)胞活力檢測(cè)的專家
靈敏、簡(jiǎn)便、低毒
關(guān)鍵詞:細(xì)胞增殖;細(xì)胞毒性;CCK-8;MTT產(chǎn)品背景
細(xì)胞功能實(shí)驗(yàn)中,對(duì)于細(xì)胞增殖、毒性和活力的檢測(cè)是至關(guān)重要的。目前常用的細(xì)胞增殖與活性檢測(cè)試劑通常是通過(guò)基于線粒體內(nèi)的脫氫還原酶的還原能力來(lái)進(jìn)行檢測(cè)的,如MTT法,XTT法,WST-1法,CCK 法等。
翊圣生物推出一款基于WST-8的細(xì)胞活性檢測(cè)試劑盒 Cell Counting Kit簡(jiǎn)稱CCK試劑盒,相較于MTT等其細(xì)胞毒性更低,檢測(cè)更靈敏,操作更簡(jiǎn)便。
實(shí)驗(yàn)原理
WST-8(2-(2-甲氧基-4-硝苯基)-3-(4-硝苯基)-5-(2,4-二磺基苯)-2H-四唑單鈉鹽)在電子耦合載體1-Methoxy PMS存在的情況下,可以被線粒體內(nèi)的脫氫酶還原生成高度水溶性的橙黃色甲臜產(chǎn)物(formazan),顏色的深淺與細(xì)胞的增殖成正比,與細(xì)胞毒性成反比。使用酶標(biāo)儀在450nm波長(zhǎng)處測(cè)定OD值,間接反映活細(xì)胞數(shù)量。
表1 CCK法與其他細(xì)胞增殖/毒性檢測(cè)方法的優(yōu)勢(shì)比較
檢測(cè)方法
MTT法
XTT法
WST-1法
CCK 法
甲臜產(chǎn)物的水溶性
差(需加有機(jī)溶劑溶解檢測(cè))
好
好
好
產(chǎn)品性狀
粉末
2瓶溶液
溶液
1瓶溶液
使用方法
配成溶液后使用
現(xiàn)配現(xiàn)用
即開(kāi)即用
即開(kāi)即用
檢測(cè)靈敏度
高
很高
很高
高
檢測(cè)時(shí)間
較長(zhǎng)
較短
較短
zui短
檢測(cè)波長(zhǎng)
560-600 nm
420-480 nm
420-480 nm
430-490 nm
細(xì)胞毒性
高,細(xì)胞形態(tài)完全消失
很低,細(xì)胞形態(tài)不變
很低,細(xì)胞形態(tài)不變
很低,細(xì)胞形態(tài)不變
試劑穩(wěn)定性
一般
較差
一般
很好
批量樣品檢測(cè)
可以
非常適合
非常適合
非常適合
便捷程度
一般
便捷
便捷
非常便捷
產(chǎn)品特點(diǎn)操作簡(jiǎn)單:產(chǎn)品本身是液體,即開(kāi)即用;甲臜產(chǎn)物是水溶性的,不需要用有機(jī)溶劑溶解。
靈敏度高:線性范圍廣,靈敏度高,重復(fù)性好。
細(xì)胞毒性低:細(xì)胞毒性非常低,對(duì)后續(xù)實(shí)驗(yàn)沒(méi)有影響。
穩(wěn)定性好:培養(yǎng)液中酚紅和血清的存在不會(huì)對(duì)檢測(cè)造成影響。
應(yīng)用性廣:可用于大規(guī)模、高通量樣品檢測(cè)。
適用范圍細(xì)胞增殖測(cè)定、細(xì)胞毒性、藥物篩選、腫瘤藥敏試驗(yàn)、生物因子的活性檢測(cè)等
產(chǎn)品數(shù)據(jù)同類產(chǎn)品性能比較:
細(xì)胞類型:293T人胚腎細(xì)胞;培養(yǎng)基:10% FBS 高糖DMEM培養(yǎng)基
孵育時(shí)間:3h;細(xì)胞數(shù)目:1000
引用已發(fā)表文獻(xiàn)數(shù)據(jù)GIST-T1細(xì)胞增殖檢測(cè)
CRTL為空白對(duì)照組,Lv-shNC為陰性干擾組,Lv-shOrai1為Orai1敲除組
ROS17/2.8細(xì)胞增殖檢測(cè)
WT為野生型,KO為BK基因敲除組
rMSCs細(xì)胞活性檢測(cè)
在2.5,5,10,20ug/mL的濃度 G/SWCNT hybrids, G, and SWCNTs處理下的細(xì)胞活性檢測(cè)
rMSCs細(xì)胞增殖檢測(cè)
G/SWCNT hybrids 在2.5,5,10,20ug/mL的濃度下處理1,3和7天細(xì)胞活性檢測(cè)
Huh6細(xì)胞干擾EZH2基因和陰性對(duì)照組細(xì)胞活性檢測(cè)
HepG2細(xì)胞干擾EZH2基因和陰性對(duì)照組細(xì)胞活性檢測(cè)
GIST-T1 細(xì)胞活性檢測(cè)
分別在2-APB 不同濃度0, 50, 100, and 150 M處理下的細(xì)胞活力檢測(cè)
GIST-T1 細(xì)胞活性檢測(cè)
分別在SKF-96365不同濃度下
0, 0.5, 1.0, and 1.5 M處理下胞活力檢測(cè)
FAQQ1:CCK的保存和穩(wěn)定性如何?
答:CCK穩(wěn)定性高,避光條件-20℃有效期2年,4℃有效期1年。經(jīng)常使用建議4℃保存,避免反復(fù)凍融。CCK正常顏色為粉紅色,若顏色發(fā)生明顯偏差,質(zhì)量可能有發(fā)生變化。
Q2:CCK的細(xì)胞毒性如何?
答:CCK毒性非常低,因此CCK檢測(cè)完后相同的細(xì)胞還能用于其它細(xì)胞增殖檢測(cè)如結(jié)晶紫染色法,中性紅染色法或DNA熒光染色法。
Q3:CCK會(huì)對(duì)活細(xì)胞進(jìn)行染色嗎?
答:不會(huì),首先WST-8不會(huì)進(jìn)入細(xì)胞染色,整個(gè)顯色反應(yīng)都是在培養(yǎng)體系中進(jìn)行的。另外WST和WST-8甲臜都屬于高度水溶性組分,反應(yīng)結(jié)束更換新的培養(yǎng)液,即可清除外源組分。
Q4:做細(xì)胞活性檢測(cè)時(shí),每孔應(yīng)該接種多少個(gè)細(xì)胞?
答:當(dāng)使用標(biāo)準(zhǔn)96孔板時(shí),貼壁細(xì)胞的zui小接種量至少為1,000個(gè)/孔(100 L培養(yǎng)基)。檢測(cè)白細(xì)胞時(shí)的靈敏度相對(duì)較低,因此推薦接種量不低于2,500個(gè)/孔(100 L培養(yǎng)基)。
Q5:若是使用6孔或者24孔板進(jìn)行試驗(yàn),CCK試劑使用量怎么樣呢?
答:如果要使用6孔板或24孔板實(shí)驗(yàn),請(qǐng)先計(jì)算每孔相應(yīng)的接種量,并按照每孔培養(yǎng)基總體積的10%加入CCK溶液。
Q6:培養(yǎng)基的本底顏色如酚紅會(huì)不會(huì)影響細(xì)胞活性的檢測(cè)呢?
答:不會(huì)的,培養(yǎng)基中酚紅的吸光度可以在計(jì)算時(shí),通過(guò)扣除空白孔中本底的吸光值而消去,因此不會(huì)對(duì)檢測(cè)造成影響。
Q7:能否用384孔板進(jìn)行細(xì)胞增殖與活性檢測(cè)的實(shí)驗(yàn)?zāi)兀?/p>
答:可以,CCK-8產(chǎn)品的使用量為每孔培養(yǎng)基總體積的10%。建議先將CCK-8產(chǎn)品稀釋一倍,然后加入培養(yǎng)基總體積的20%即可,這樣可減少誤差。
Q8:只可以在450 nm波長(zhǎng)處測(cè)OD值嗎?
答:可以在450 nm波長(zhǎng)處測(cè)OD值,但是若無(wú)此波長(zhǎng)的濾光片,可選擇吸光度在430-490 nm之間的濾光片,但是450 nm濾光片的檢測(cè)靈敏度zui高。
Q9:若是暫不檢測(cè)OD值,該如何處理?
答:若暫時(shí)不測(cè)定OD值,可以向每孔中加入10 L 0.1 M的HCL溶液或者1% w/v SDS溶液,并遮蓋培養(yǎng)板避光保存在室溫條件下。24小時(shí)內(nèi)測(cè)定,吸光度不會(huì)發(fā)生變化
Q10:在加入CCK-8時(shí)可否不更換培養(yǎng)基?
答:一般情況下可以不更換培養(yǎng)基。但是如果培養(yǎng)基里有氧化還原性物質(zhì)的話,有可能會(huì)產(chǎn)生誤差,必須更換其它培養(yǎng)基。
Q11:在實(shí)驗(yàn)中OD值太高,怎么解決?
答:可以縮短加入CCK-8后的培養(yǎng)時(shí)間。例如:可以把加入CCK-8試劑后的培養(yǎng)時(shí)間由2小時(shí)縮短為1小時(shí)。此外,可適當(dāng)減少細(xì)胞的數(shù)量。
Q12:如果OD值太低,怎么解決?
答:可以采取2個(gè)辦法:1、適當(dāng)增加細(xì)胞數(shù)量 2、延長(zhǎng)加入CCK-8試劑后的反應(yīng)時(shí)間。
Q13:應(yīng)該每次做標(biāo)準(zhǔn)曲線嗎?
答:建議每次做。雖然細(xì)胞是一樣的,但是細(xì)胞的狀態(tài)不一定一樣。對(duì)于狀態(tài)不一樣的細(xì)胞,建議每次做標(biāo)準(zhǔn)曲線。
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產(chǎn)品信息
產(chǎn)品名稱
產(chǎn)品編號(hào)
規(guī)格
價(jià)格(元)
促銷價(jià)(元)
Cell Counting Kit(CCK-8)CCK-8試劑盒
40203ES60
100T (1 mL)
150.00
150.00
Cell Counting Kit(CCK-8)CCK-8試劑盒
40203ES76
500T (5 mL)
480.00
480.00
Cell Counting Kit(CCK-8)CCK-8試劑盒
40203ES80
1000T (10 mL)
920.00
897.00
Cell Counting Kit(CCK-8)CCK-8試劑盒
40203ES88
3000T (30 mL)
1833.00
1687.00
Cell Counting Kit(CCK-8)CCK-8試劑盒
40203ES92
10000T (100 mL)
5122.00
4897.00
更多產(chǎn)品敬請(qǐng)?jiān)斣?/p>