RIPA Buffer (strong)
Cat.No:R0010 Solarbio
Storage:Store PMSF at -20℃, lysis buffer at 2-8℃, valid for 1 year.
Appearance:Yellow clarified liquid
Qty:
Size:
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RIPA Buffer (strong)Storage:Store PMSF at -20℃, lysis buffer at 2-8℃, valid for 1 year.
Appearance:Yellow clarified liquid
Qty:
Size:
Name | RIPA buffer(high) |
Appearance | Yellow clarified liquid |
Storage | Store PMSF at -20℃, lysis buffer at 2-8℃, valid for 1 year. |
Unit | Bottle |
Specification | 20ml 100ml 500ml |
Transport temperature: normal temperature transport
This product is a highly efficient tissue cell lysate. It is mainly used to extract soluble proteins from animal tissues and animal cells. Tissue Live cells This product is equipped with a PMSF (0.3ml/1.5ml), the PMSF should be stored at -20℃.
Instructions for use
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, 10ul PMSF is added to every 1ml RIPA to make the final concentration of PMSF 1mM. Mix well and set aside (PMSF add 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: Collect the cells centrifugally and use your fingers to forcefully flick the cells apart. Add the lysate at the ratio of 150-250 ul per cell volume of the 6-well plate, and then flick with your finger to fully lysate the cells. There should be no obvious cell precipitation after full lysis. If the number of cells is large, it must be divided into 500,000 to 1 million cells/tubes, and then cracked.
c) For tissue samples: Cut the tissue into fine fragments. The lysate is added at a ratio of 150-250 ul lysate 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:
This reagent is a strong lysate, which can extract nuclear protein, but at the same time of extracting nuclear protein, the genome will also be released, resulting in a thick cell lysate. At this time, the protein sample buffer can be directly added, boiled and centrifuged, and the sample is directly loaded after centrifugation. To determine the concentration, add a small amount of SDS (1%), boil and centrifuge the concentration. The protein extracted by this series of protein extraction reagents contains stain remover, so it is not suitable for Bradford protein concentration determination kit. Please choose BCA method or Lowry method to detect protein concentration.
If a protein that is particularly tightly bound to the genome needs to be detected, the goo can be broken up by ultrasound treatment, then the supernatant can be centrifuged for subsequent experiments.
Note:Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.
Remark:These protocols are for reference only. Solarbio does not independently validate these methods.
Note:
1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.
2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.
3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.
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请问R0010高效型裂解液里,有磷酸酶抑制剂吗? 2023-07-16 22:28:13
Administrator您好,裂解液中不包含磷酸酶抑制剂,只搭配一支PMSF,有需要的需要自行添加磷酸酶抑制剂
裂解液中含SDS吗 2023-07-12 15:19:43
Administrator您好,裂解液中含有SDS
高效RIPA裂解液能提取到膜蛋白吗?能的话膜蛋白在离心后的上清里吗? 2023-06-01 17:08:22
Administrator您好,高效RIPA裂解液是全蛋白提取试剂盒,能提到膜蛋白,离心后在上清中,后续实验仅针对膜蛋白的话可能效果不太理想,杂蛋白偏多。仅提膜蛋白的话本公司有专门针对动植物组织和细胞,细菌,真菌,昆虫等膜蛋白提取的试剂盒。
请问加了RIPA以后,没有加PMSF,加了其他的蛋白酶抑制剂可以吗?官网没有说裂解液里有磷酸酶抑制剂啊 2022-09-01 17:30:58
Administrator可以加其他蛋白酶抑制剂,裂解液里不含磷酸酶抑制剂,需要根据需求自己添加
请问加强型ripa裂解液中,是否有磷酸酶抑制剂和蛋白酶抑制剂呢,没看到说明书又说,但是使用说明中并未提及要单独补充。 2022-07-25 19:06:21
Administrator官网有展示说明书,裂解液里面含有磷酸酶抑制剂,随带一支蛋白酶抑制剂PMSF.