Synonyms | 蛋白提取试剂盒 |
English Name | RIPA buffer |
Chinese Name | 普通RIPA裂解液(组织/细胞) |
Storage | Store PMSF at -20℃, lysis buffer at 2-8℃, valid for 1 year. |
Unit | Bottle |
Specification | 100ml 500ml |
Product Description: RIPA tissue/cell lysate is the use of surfactants, etc., to cleave cell membranes (including nuclear membranes). The protein extracted by this reagent is soluble total protein.
Instruction:
If RIPA is found to have precipitation, please let it sit at room temperature for half an hour or in a room temperature water bath to dissolve the precipitation.
According to the amount used, add 10ul PMSF for every 1ml RIPA, so that the final concentration of PMSF is 1mM, mix and set aside (add PMSF now).
1. Sample pretreatment:
a) For adherent cells: Remove the culture medium and wash it again with PBS, normal saline or serum-free culture medium. The lysate was added at the ratio of 150-250 ul lysate per cell volume of the 6-well plate. Blow the gun a few times to make full contact between the lysate and the cells.
b) For suspended cells: Centrifuge the cells and add 150-250ul of lysate according to the number of cells in each well of the 6-well plate. Tap the suspended cells with your fingers to fully lysate them. There should be no obvious cell precipitation after full lysis. If the number of cells is large, it must be divided into 5-10×105 cells/tubes, and then cracked.
c) For tissue samples: Cut the tissue into fine fragments. Add the lysate at a ratio of 150-250ul per 20mg of tissue
(If the cleavage is insufficient, more lysate can be appropriately added, and if a high concentration of protein samples is required, the amount of lysate can be appropriately reduced).
Homogenize with a glass homogenizer until fully cracked.
2. Post-processing:
Centrifuge 10000-14000g of the cracked sample for 3-5 minutes, take the supernatant, and then perform subsequent PAGE, Western and immunoprecipitation operations.
Note:Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.
请问里面含有磷酸酶抑制剂吗
2023-06-02 12:49:42
Administrator您好,不含磷酸酶抑制剂。
这个裂解液里含有SDS.嘛2023-05-29 17:13:00
Administrator您好,有SDS成分。
可以用于Co-IP吗?2023-05-14 14:40:37
Administrator您好,提取的蛋白是变性蛋白,不可以用于Co-IP
这个可以裂解外泌体蛋白吗?使用比例麻烦告知2023-03-12 14:17:25
Administrator理论上是可以的,按照外泌体和RIPA裂解液比例1:1裂解,或者也可以参考RIPA裂解液说明书中细胞裂解的方法。
请问能否提取膜蛋白,后续能否做ELISA2022-10-04 21:14:50
Administrator普通型RIPA裂解液对膜蛋白提取效率较低。如需提取膜蛋白,建议依据样本种类可在索莱宝官网搜索专用的膜蛋白提取试剂盒,并查看相关说明书。