MTT细胞增殖及细胞毒性检测试剂盒
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产品编号 C0009M
数量
¥ 759.00
产品包装:
500次 2500次
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使用说明
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MTT细胞增殖及细胞毒性检测试剂盒(MTT Cell Proliferation and Cytotoxicity Assay Kit)是一种非常经典的细胞增殖和细胞毒性检测试剂盒,被广泛应用于细胞增殖和细胞毒性的检测。
MTT可以被线粒体内的一些脱氢酶还原生成结晶状的深紫色产物formazan(图1A)。在特定溶剂存在的情况下,可以被完全溶解(图1B)。然后通过酶标仪可以测定570nm波长附近的吸光度(图2)。细胞增殖越多越快,则吸光度越高;细胞毒性越大,则吸光度越低。

图1.MTT细胞增殖及细胞毒性检测试剂盒实测效果图。A.HeLa细胞加入使用本试剂盒配制的MTT溶液,在细胞培养箱内孵育4小时,显微镜下可见大量结晶状的深紫色产物formazan生成。B.不同数量HeLa细胞在MTT溶液(MTT solution)加入后4小时的效果图(上图)及深紫色产物formazan生成后加入Formazan溶解液(Formazan solvent),充分溶解后的效果图(下图)。

图2. 本试剂盒检测不同数量HeLa细胞的效果图。不同的检测条件下,实际读数会因标准品的配制、检测仪器等的不同而存在差异,图中数据仅供参考。

本试剂盒采用了独特的Formazan 溶解液配方,无需去除原有的培养液,可以直接加入Formazan溶解液溶解formazan。从而避免了由于去除培养液时formazan被部分去除而引起的误差。
本试剂盒本底低,灵敏度高,线性范围宽,使用方便。
碧云天各种细胞增殖和细胞毒性检测试剂盒的比较和选择,请参考 http://www.beyotime.com/cell-proliferation.htm
本试剂盒C0009S包装可以测定500个样品,C0009M包装可以测定2500个样品。
包装清单:

产品编号 产品名称 包装
C0009S-1 MTT 25mg
C0009S-2 MTT溶剂 5ml
C0009S-3 Formazan溶解液 55ml
说明书 1份

产品编号 产品名称 包装
C0009M-1 MTT 125mg
C0009M-2 MTT溶剂 25ml
C0009M-3 Formazan溶解液 250ml
说明书 1份

保存条件:
4℃或-20℃保存,MTT需避光保存,一年有效;MTT配制成溶液后需-20℃避光保存;Formazan溶解液也可以室温保存。
注意事项:
由于使用96孔板进行检测,如果细胞培养时间较长,一定要注意蒸发的问 题。一方面,由于96孔板周围一圈最容易蒸发,可以采取弃用周围一圈的办法,改加PBS,水或培养液;另一方面 ,可以把96孔板置于靠近培养箱内水源的地方,以缓解蒸发。
MTT溶剂在低温情况下会凝固,使用前请室温放置或20-25℃水浴温育片刻 至全部融解后使用。
MTT配制成溶液后为黄色,需避光保存,长时间光照会导致失效。当颜色变为灰绿色时,请勿使用。MTT溶液在4℃、冰浴等较低温度情况下会凝固,可以20-25℃水浴温育片刻至全部融解 后使用。
Formazan溶解液结冻或产生沉淀时可以室温或37℃水浴孵育以促进溶解,并且必须在全部溶解并混匀后使用。
加入Formazan溶解液后请适当轻轻混匀,但须注意避免产生泡沫。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1. MTT溶液的配制:用5ml MTT溶剂溶解25mg MTT,配制成5mg/ml的MTT溶液。配制后即可使用,或直接-20℃避光保存,也可以根据需要适当分装后-20℃避光保存。
2. 通常细胞增殖实验每孔加入100微升2000个细胞,细胞毒性实验每孔加入100微升5000个细胞(具体每孔所用的细胞的数目,需根据细胞的大小,细胞增殖速度的快慢等决定)。按照实验需要,进行培养并给予0-10微升特定的药物刺激。
3. 每孔加入10微升MTT溶液,在细胞培养箱内继续孵育4小时。
4. 每孔加入100微升Formazan溶解液,适当混匀,在细胞培养箱内再继续孵育。直至在普通光学显微镜下观察发现formazan全部溶解。通常37℃孵育3-4小时左右,紫色结晶会全部溶解。如果紫色结晶较小较少,溶解的时间会短一些。如果紫色结晶较大较多,溶解的时间会长一些,此时为加速溶解可以适当振摇数次。
5. 在570nm测定吸光度(可参考图2)。如无570nm滤光片,可以使用560-600nm的滤光片。



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