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    普利莱
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    C1053-100
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描述: RIPA裂解液(RIPA Lysis Buffer)对动物细胞胞膜、胞浆、胞核成分均有较强裂解作用,是经典最常用的细胞组织快速裂解液。使用RIPA 裂解缓冲液制备用于Western特别是用于免疫共沉淀的裂解产物已经是一种首选的标准操作,适用于大部分抗原表位检测,特别免疫共沉淀等实验

规格:100ml  500ml

成分:50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% NP-40, 0.1% SDS.

储存:4 ℃保存 12个月有效

使用说明:

制备细胞裂解产物

1. 800g 4℃离心5分钟收集培养细胞,估计细胞离心后的体积(PCV, 106 cells=~20ul, 107 cells =~100ul PCV);

2. 50100ul PCV加入5倍体积RIPA裂解缓冲液(250~500ul,冰浴放置10分钟,并每隔5分钟在漩涡混合仪上振荡30秒;

3. 12000g 4℃离心10分钟,将上清转移到新的离心管中,即得细胞总蛋白产物。

注意:如所得蛋白产物较为粘稠,可先95℃加热5分钟,迅速冰浴5分钟,再进行步骤3

制备组织裂解产物

1. 50-100mg组织在冰上剪成碎片,用预冷的PBS洗涤2次离心弃去PBS

2. 加入 0.5-1ml 预冷 RIPA 裂解缓冲液;

3. 4℃用玻璃匀浆器匀浆20-40次,直到95%的细胞被破碎,然后在冰浴中放置10分钟,并每隔5分钟在漩涡混合仪上振荡30秒;

4. 12000g 4℃离心10分钟,将上清转移到新的离心管中,即得组织总蛋白产物。


注意:如所得蛋白产物较为粘稠,可先95℃加热5分钟,迅速冰浴5分钟,再进行步骤4

说明:

1. 转移上清液时不要吸入底部的沉淀物;

2. 在做免疫沉淀或免疫共沉淀时最好在实验前进行蛋白提取,避免某些不稳定蛋白的降解;

3. RIPA裂解缓冲液中未加入蛋白酶抑制剂,用户可自行选择添加。

使用我司RIPA裂解缓冲液发表SCI论文已达数百篇,部分文章列表如下,供参考:

1、Corporation H P. Secernin-1 Contributes to Colon Cancer Progression through Enhancing Matrix Metalloproteinase-2/9 Exocytosis.[J]. Disease Markers, 2015.

2、Corporation H P. Cytotoxic Effects of Biosynthesized Zinc Oxide Nanoparticles on Murine Cell Lines[J]. Evidence-based Complementary and Alternative Medicine : ECAM, 2015, 2015.

3、Corporation H P. Protective effects of N-acetylcysteine in concanavalin a-induced hepatitis in mice.[J]. Mediators Inflamm, 2015.

4、Corporation H P. Blockade of lysosomal acid ceramidase induces GluN2B-dependent Tau phosphorylation in rat hippocampal slices.[J]. Neural Plasticity, 2014,

5、J. V, T. P, Y. L, et al. Androgen Receptor Activation[J]. Cellular Physiology & Biochemistry, 2014, 33(6):1911-1920.

6、Yang K, Song Y, Tang Y, et al. mAChRs activation induces epithelial-mesenchymal transition on lung epithelial cells.[J]. Bmc Pulmonary Medicine, 2014, 14(4):417-419.

7、Peng Y, Guo J, Liu Y, et al. MicroRNA-34A inhibits the growth, invasion and metastasis of gastric cancer by targeting PDGFR and MET expression[J]. Bioscience Reports, 2014, 34(2):247-256.

8、Lee S Y. Nonthermal plasma induces apoptosis in ATC cells: involvement of JNK and p38 MAPK-dependent ROS.[J]. Yonsei Medical Journal, 2014, 55(6).

9、Jeong J, Conboy M J, Conboy I M. Pharmacological inhibition of myostatin/TGF-β receptor/pSmad3 signaling rescues muscle regenerative responses in mouse model of type 1 diabetes[J]. Acta Pharmacologica Sinica, 2013, 34:1052-1060.

10、Vaziri N D, Yuan J, Nazertehrani S, et al. Chronic Kidney Disease Causes Disruption of Gastric and Small Intestinal Epithelial Tight Junction.[J]. American Journal of Nephrology, 2013, 38(2):99-103.

11、Li Y, Sun Y, Liu F, et al. Norcantharidin Inhibits Renal Interstitial Fibrosis by Blocking the Tubular Epithelial-Mesenchymal Transition[J]. Plos One, 2013, 8(6):e66356.

12、Kashyap M, Kawamorita N, Tyagi V, et al. Down-regulation of nerve growth factor expression in the bladder by antisense oligonucleotides as new treatment for overactive bladder.[J]. Journal of Urology, 2013, 190(2):757-764.

13、Tisdell, Francis E, et al. "3-(substituted phenyl)-5-(substituted cyclopropyl)-1,2,4-triazole compounds." US, US 6262099 B1. 2001.

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