基本描述

产品名称	植物RIPA裂解液(强)
产品介绍	阿拉丁生产的植物RIPA裂解液(Plant RIPA Lysis Buffer)是一种传统的植物细胞、组织或原生质体快速裂解液。植物RIPA裂解液裂解得到的蛋白样品可以用于常规的PAGE、Western、免疫沉淀(immunol precipitation，IP)、免疫共沉淀(co-IP)和ELISA等。本产品可以用于植物细胞、组织或原生质体样品，也可以用于动物的细胞或组织样品、真菌或细菌样品。 RIPA的本意是Radio Immunoprecipitation Assay。关于不同的RIPA裂解液以及阿拉丁生产的其它裂解液的主要特点和差异，以及如何选择裂解液可参考我们的相关网页：http://www.aladdin-e.com/。植物RIPA裂解液(强)的主要有效成分包括1% Triton X-100、1% sodium deoxycholate和0.1% SDS等去垢剂，以及sodium orthovanadate、sodium fluoride、EDTA和leupeptin等多种抑制剂。可以有效抑制蛋白降解。使用说明： 1.对于培养的植物细胞样品： a.融解植物RIPA裂解液，混匀。取适量的裂解液，在使用前数分钟内加入PMSF，使PMSF的最终浓度为1mM，或者根据实验需要加入适当的上述蛋白酶磷酸酶抑制剂混合物。 b.离心收集植物细胞，按照每50-100万细胞加入100-200微升裂解液的比例加入裂解液。用枪吹打数下，使裂解液和细胞充分接触，在冰上裂解2-10分钟。 c.充分裂解后，10000-14000g离心3-5分钟，取上清，即可进行后续的PAGE、Western和免疫沉淀等操作。 2.对于植物原生质体样品： a.把组织剪切成细小的碎片，制备原生质体。 b.(可选)质粒转化原生质体，继续培养16-48小时，并可以根据需要给予适当的实验条件处理。 c.100-500g低速离心收集原生质体。 d.融解植物RIPA裂解液，混匀。取适量的裂解液，在使用前数分钟内加入PMSF，使PMSF的最终浓度为1mM，或者根据实验需要加入适当的蛋白酶磷酸酶抑制剂混合物。 e.按照每50-100万原生质体加入100-200微升裂解液的比例加入裂解液。轻弹管底以充分裂解细胞。充分裂解后应没有明显的原生质体沉淀。如果原生质体量较多，必需分装成50-100万原生质体/管，然后再裂解。大团的50-100万原生质体较难裂解充分，而少量的50-100万原生质体由于裂解液容易和50-100万原生质体充分接触，相对比较容易裂解充分。 3.对于植物组织样品： a.把植物组织剪切成细小的碎片。 b.融解植物RIPA细胞裂解液(强)，混匀。取适量的裂解液，在使用前数分钟内加入PMSF，使PMSF的最终浓度为1mM，或者根据实验需要加入适当的蛋白酶磷酸酶抑制剂混合物。 c.按照每20毫克植物组织加入100-200微升裂解液的比例加入裂解液。(如果裂解不充分可以适当添加更多的裂解液，如果需要高浓度的蛋白样品，可以适当减少裂解液的用量。) d.用玻璃匀浆器匀浆，或使用阿拉丁生产的E6600 TissueMaster™手持式组织研磨仪研磨，直至充分裂解。也可以把组织样品冷冻后液氮研磨，研磨充分后加入裂解液进行裂解。 e.充分裂解后，10000-14000g离心3-5分钟，取上清，即可进行后续的PAGE、Western和免疫沉淀等操作。 f.如果植物组织样品本身非常细小和柔嫩，可以适当剪切后直接加入裂解液裂解，通过强烈vortex使样品裂解充分。然后同样离心取上清，用于后续实验。直接裂解的优点是比较方便，不必使用匀浆器或研磨设备，缺点是不如匀浆或研磨那样裂解比较充分。注：植物RIPA裂解液的裂解产物中经常会出现一小团透明胶状物，属正常现象。该透明胶状物为含有基因组DNA等的复合物。在不检测和基因组DNA结合特别紧密的蛋白的情况下，可以直接离心取上清用于后续实验；如果需要检测和基因组结合特别紧密的蛋白，则可以通过超声处理打碎打散该透明胶状物，随后离心取上清用于后续实验。如果检测一些常见的转录因子，通常不必进行超声处理，就可以检测到这些转录因子。 PMSF (ST506) is required, but not supplied in this product. Inhibitor cocktails can also be used for better lysis results. provides a variety of inhibitor cocktails, such as Protease and Phosphatase Inhibitor Cocktail for General Use , Protease and Phosphatase Inhibitor Cocktail for General Use, MS-safe , Protease and Phosphatase Inhibitor Cocktail for Mammals , Protease and Phosphatase Inhibitor Cocktail for plant , Protease and Phosphatase Inhibitor Cocktail for Fungi , and Protease and Phosphatase Inhibitor Cocktail for Bacteria . If phosphorylated proteins are not to be detected, inhibitors of protease only can be used.All procedures for sample lysis should be performed on ice or at 4℃. An appropriate sample lysis buffer can be selected by referring to the website at https://www.aladdin-e.com or optimized by preliminary tests. This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use： 1. Lysis of cultured cells a. Thaw Plant RIPA Lysis Buffer completely and mix well. Just before use, take an appropriate amount of RIPA Lysis Buffer and add PMSF to a final concentration of 1mM. Other protease and phosphatase inhibitors can also be used if required. b. Collect cells by centrifugation, add 100-200μl of lysis buffer per 0.5-1.0×106 cells. Pipette to resuspend cells and lyse cells on ice for 2-10 min. c. After full lysis of cells, centrifuge at 10,000-14,000×g for 3-5 minutes and take the supernatant for PAGE, Western or IP analysis. 2. Lysis of protoplasts a. Cut tissues into small pieces and prepare protoplasts. b. (Optional) Transform the plasmid into protoplasts and continue incubation for 16-48 hours. Treat as desired. c. Centrifuge at 100-500×g to collect protoplasts. d. Thaw Plant RIPA Lysis Buffer completely and mix well. Just before use, take an appropriate amount of Lysis Buffer and add PMSF to a final concentration of 1mM. Other protease and phosphatase inhibitors can also be used if required. e. Add 100-200μl of lysis buffer per 0.5-1.0×106 protoplasts, flick the bottom of the tube to lyse thoroughly. There should be no obvious precipitates after full lysis. For large amounts of protoplasts, dispense them into 0.5-1.0×106 protoplasts per tube for lysis. 3. Lysis of tissues a. Cut tissues into small pieces. b. Thaw Plant RIPA Lysis Buffer completely and mix well. Just before use, take an appropriate amount of Lysis Buffer and add PMSF to a final concentration of 1mM. Other protease and phosphatase inhibitors can also be used if required. c. Add 100-200μl of lysis buffer per 20mg of tissues. The amount of lysis buffer can be adjusted based on the lysis results. d. Homogenize tissues thoroughly with a glass homogenizer or 's TissueMasterTM Handheld Homogenizer . e. Centrifuge at 10,000-14,000×g for 3-5 minutes and take the supernatant for PAGE, Western or IP analysis. f. For tiny tissues, add lysis buffer after appropriate cutting and lyse tissues with vigorous vortex. Centrifuge and take the supernatant for subsequent analysis. This lysis method is convenient, with no homogenization needed, but the lysis result is not as thorough as that from the homogenization method. Note: It is normal to see a small clump of transparent gelatinous substances containing genomic DNA in RIPA cell lysate. In cases where DNA-binding proteins will not be analyzed, the supernatant can be analyzed directly. Otherwise, break the gelatinous substance with an ultrasonicator and analyze the supernatant after centrifugation. Some common transcription factors such as NF-kappaB and p53 can be detected in the supernatant without ultrasonic processing

储存与运输

储存温度	-20°C储存
运输条件	超低温冰袋运输
稳定性与储存	-20℃保存，一年有效。

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植物RIPA裂解液(强)

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