1. MCE Kits
  2. Protein Biology
  3. Protein Sample Preparation
  4. Cell Lysis
  5. RIPA Lysis Buffer (Strong)

RIPA Lysis Buffer (Strong) 

RIPA 裂解液 (强)

Cat. No.: HY-K1001
Manual SDS

RIPA Lysis Buffer (Strong) 是一种传统的细胞、组织快速裂解液,适用于 Western Blot (WB)、免疫沉淀 (IP) 及免疫共沉淀 (Co-IP) 等实验。

RIPA Lysis Buffer (Strong)

1.  客户无需承担相应的运输费用。

2.  同一机构(单位)同一产品试用装仅限申领一次,同一机构(单位)一年内

     可免费申领三个不同产品的试用装。

3.  试用装只面向终端客户

规格 价格 是否有货 数量
Free Sample (10 mL)   Apply now  
100 mL ¥395 In-stock

* Please select Quantity before adding items.

注册 MCE会员完成审核
即刻享有 积分商城 300 专属积分

  • Description

  • Storage

  • Protocol

  • Documentation

Description
& Advantages

MCE RIPA (Radio-Immunoprecipitation Assay) Lysis Buffer (Strong) is a widely used lysis and wash buffer for reporter assays, protein kinase assays, immunoassays and protein purification.

RIPA Lysis Buffer (Strong) consists of 50 mM Tris (pH 7.4), 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, and general protease and phosphatase inhibitors (e.g., sodium orthovanadate, sodium fluoride, EDTA). If desired, add MCE Protease Inhibitor Cocktail (HY-K0010) and MCE Phosphatase Inhibitor Cocktails (e.g., HY-K0021, HY-K0022, and HY-K0023).

Publications
Storage

Stored at -20°C, and is stable for up to 12 months.

Protocol

1.   Procedure for Cultured Cells

1.1   Pipette proper volume of RIPA Lysis Buffer and mix thoroughly. Add Protease Inhibitor Cocktail to lysis buffer to prevent proteolysis. Add Phosphatase Inhibitor Cocktails to maintain phosphorylation status of proteins as needed. Incubate on ice for subsequent use.

1.2   For adherent cells: Carefully remove culture medium from adherent cells. Wash cells twice with cold wash solution, such as PBS, normal saline or serum-free culture medium, to remove residual medium. Add proper volume of cold RIPA Lysis Buffer, then stroke with pipette until the buffer immerses cells completely. Incubate on ice and shake slightly for 5-10 minutes.

For suspension cultured cells: Collect cells into a centrifuge tube. Centrifuge samples at 500 g for 5 minutes and discard the supernatants. Wash cells twice with cold wash solution, such as PBS, normal saline or serum-free culture medium, to remove residual medium. Add proper volume of cold RIPA Lysis Buffer, then stroke with pipette until the buffer immerses cells completely. Incubate on ice and shake slightly for 5-10 minutes.

Note: Generally, use 1 mL of RIPA Lysis Buffer for 0.5-5 × 106 cells. The volume of added RIPA Lysis Buffercan be adjusted proportionally.

1.3   After lysis, centrifuge at 14,000 g for 5 minutes at 4°C. Transfer the supernatants to a new tube for further analysis. Aliquot and freeze in liquid nitrogen and stored at –80°C for future use. Avoid multiple freeze/thaw cycles.

2.   Procedure for Tissue Lysis

2.1   Pipette proper volume of RIPA Lysis Buffer and mix thoroughly. Add Protease Inhibitor Cocktail to lysis buffer to prevent proteolysis. Add Phosphatase Inhibitor Cocktails to maintain phosphorylation status of proteins as needed. Incubate on ice for subsequent use.

2.2   Cut tissue sample into small pieces on ice quickly to minimize protein degradation.

2.3   Add 150-250 μL cold RIPA Lysis Buffer per 20 mg of tissue sample and homogenize using electric homogenizer. Add more Lysis buffer if tissue is not completely lysed.

2.4   Transfer complete homogenized sample into a centrifuge tube. Incubate on ice and shake slightly for 5-10 minutes.

2.5   After lysis, centrifuge at 14,000 g for 5 minutes at 4°C. Transfer the supernatants to a new tube for further analysis. Aliquot and freeze in liquid nitrogen and stored at –80°C for future use. Avoid multiple freeze/thaw cycles.

Note: 20 mg of frozen mouse liver tissues may yield 15-25 mg/mL protein.

Documentation
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

您最近查看的产品:

Your information is safe with us. * Required Fields.

   产品名称:

 

* 需求量:

* 客户姓名:

 

* Email:

* 电话:

 

* 公司或机构名称:

   留言给我们:

Bulk Inquiry

Inquiry Information

产品名称:
RIPA 裂解液 (强)
目录号:
HY-K1001
需求量: