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线粒体和胞浆蛋白制备试剂盒(组织和细胞通用) C1260
  • 品  牌:

    普利莱
  • 货  号:

    C1260-50
  • 规格:

  • 购买数量:

    +
  • 价格:560.00-980.00
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描述:线粒体和胞浆蛋白制备试剂盒 (Mitochondria Isolation Kit)用于从组织或培养细胞中分离线粒体和细胞胞浆成分。加入分离溶液,匀浆破碎组织细胞,经过数次800g12000g离心,在60分钟内即可分离出完整的线粒体和胞浆成分。制备的线粒体具有很高的生物学活性,可进行各种功能研究如酶学测定,更可用于Western Blot,2D-胶、线粒体蛋白或DNA提取、蛋白质组学等研究。严格按照说明操作,总是能制备获得高纯度线粒体。一篇方法学研究论文发现,用普利莱试剂盒制备线粒体的得率、活性、纯度优于蔗糖密度梯度离心法和Invotrogen/Pierce线粒体提取试剂盒方法。

特点:

1、 操作简单,可在一小时内完成。

2、 实验室常规设备即可,无需超速离心。

3、 制备的线粒体具有高活性,可用于后续的各类实验分析。

4、 发表的SCI文章超过百篇。

规格:

50100次

储存:

−20°C 一年有效

操作步骤:(注意:以下所有操作均在4°C进行)

1. 样品处理:组织匀浆:100~200mg新鲜组织样品,剪为0.5cm2的碎块放入玻璃匀浆器内。加入1.5mL冰预冷的MitoSolution。研磨组织20次。培养细胞匀浆:800g5min离心收集细胞。单次提取需2-5×107个细胞。加入1.5 mL冰预冷MitoSolution重悬细胞,将细胞悬液转移到小容量玻璃匀浆器内研磨细胞30次。

2. 将匀浆液转移到离心管中,800g,4ºC离心5min

3. 取上清液。再次800g,4ºC 离心5min

4. 将上清液转移到新的离心管。10,000g,4ºC离心10min。线粒体沉淀在管底。离心后的上清含胞浆成分,可收集用于对照实验。

5. 洗涤线粒体:加入0.2mLMitoSolution,轻弹管底重悬线粒体沉淀;12,000g,4ºC离心10min弃上清,线粒体沉淀在管底。

6. 重悬线粒体:可以用MitoSolution重悬线粒体沉淀,也可以用合适后续实验的自备缓冲液来裂解线粒体沉淀,具体用量是:约每100mg组织提取的线粒体用50µl,约每5×107个细胞提取的线粒体用100µl.用量请根据组织或细胞类型进行微调。

7. 裂解线粒体后,进行蛋白浓度测定。立即使用或−70ºC保存。


目前,使用我公司线粒体/胞浆蛋白制备试剂盒发表的SCI英文论文已有多百篇,以下供参考:

1Shangguan W J, Li H, Zhang Y H. Induction of G2/M phase cell cycle arrest and apoptosis by ginsenoside Rf in human osteosarcoma MG 63 cells through the mitochondrial pathway[J]. Oncology reports, 2014, 31(1): 305-313.

2Zhou X, Zhao Y, Fang Y, et al. Hes1 is upregulated by ischemic postconditioning and contributes to cardioprotection[J]. Cell biochemistry and function, 2014, 32(8): 730-736.

3Zhang P, Pan H, Wang J, et al. Telomerase activity-independent function of telomerase reverse transcriptase is involved in acrylamide-induced neuron damage[J]. Biotechnic & Histochemistry, 2014, 89(5): 327-335.

4Zhou X L, Wan L, Xu Q R, et al. Notch signaling activation contributes to cardioprotection provided by ischemic preconditioning and postconditioning[J]. J Transl Med, 2013, 11: 251.

5Zhang F, Zhang L, Sun L, et al. Effects of Fluid Shear Stress on Expression of Smac/DIABLO in Human Umbilical Vein Endothelial Cells[J]. Current Therapeutic Research, 2013, 74: 36-40.

6Zhao G, Ma H, Shen X, et al. Role of glycogen synthase kinase 3β in protective effect of propofol against hepatic ischemiareperfusion injury[J]. Journal of Surgical Research, 2013, 185(1): 388-398.

7Sun L L, Zhang L, Meng X L, et al. Effects of fluid shear stress on the expression of Omi/HtrA2 in human umbilical vein endothelial cells[J]. Molecular medicine reports, 2013, 7(1): 110-114.

8Sun L Q, Zhao J, Zhang T T, et al. Protective effects of Salvianolic acid B on Schwann cells apoptosis induced by high glucose[J]. Neurochemical research, 2012, 37(5): 996-1010.

9Zhou Q, Li Y, Jin J, et al. Lx2-32c, a novel taxane derivative, exerts anti-resistance activity by initiating intrinsic apoptosis pathway in vitro and inhibits the growth of resistant tumor in vivo[J]. Biological and Pharmaceutical Bulletin, 2012, 35(12): 2170-2179.

10Li W, Zhang J, An W. The conserved CXXC motif of hepatic stimulator substance is essential for its role in mitochondrial protection in H 2 O 2-induced cell apoptosis[J]. FEBS letters, 2010, 584(18): 3929-3935.

11Fang N X, Yao Y T, Shi C X, et al. Attenuation of ischemiareperfusion injury by sevoflurane postconditioning involves protein kinase B and glycogen synthase kinase 3 beta activation in isolated rat hearts[J]. Molecular biology reports, 2010, 37(8): 3763-3769.

12Zhao C Q, Zhang Y H, Jiang S D, et al. Both endoplasmic reticulum and mitochondria are involved in disc cell apoptosis and intervertebral disc degeneration in rats[J]. Age, 2010, 32(2): 161-177

13Yin Q, Jin P, Liu X, et al. SDF-1α inhibits hypoxia and serum deprivation-induced apoptosis in mesenchymal stem cells through PI3K/Akt and ERK1/2 signaling pathways[J]. Molecular biology reports, 2011, 38(1): 9-16.


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