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G2033-30 / 100ML Ripa Lysis Buffer Solution Weak Biochemistry Reagent

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G2033-30 / 100ML Ripa Lysis Buffer Solution Weak Biochemistry Reagent

RIPA Lysis Buffer (RIPA Lysis Buffer) is a traditional cell and tissue rapid lysis Buffer. The protein sample after lysis with RIPA lysis Buffer can be used for regular Western, IP, etc. There are many formulas for RIPA lysis Buffer, which can be divided into strong, medium and weak according to the effect. This lysate is RIPA weak lysate, and its main components are 50 mM Tris-Hcl, 150 mM NaCl, 1 mM EDTA-Na2, 1% NP-40, 0.25% Sodium deoxycholate.

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Item code Description Volume
G2033-30ML RIPA Lysis Buffer 30mL
G2033-100ML 100mL

 

Technical Specification

Storage conditions:

Store at 4ºC in the dark, valid for 12 months.

Instructions:

For tissue samples:

  1. The tissue pieces are washed with cold PBS to remove blood stains. Cut into small pieces and place in a homogenizer.
  2. Add 10 times the volume of the tissue to this reagent (add protease inhibitor within a few minutes before use) and homogenize thoroughly with a high-speed tissue grinder.
  3. Transfer the homogenate to a 1.5 mL centrifuge tube and shake. In the ice bath for 30 minutes, pipette repeatedly to ensure complete cell lysis.
  • Centrifuge at12000g for 5 min, and collect the supernatant, which is the total protein solution.


For cultured cell samples:

1. For adherent cells:

  • A. Wash the cells 2-3 times with PBS. Thoroughly absorb the remaining liquid one last time.
  • B. Add an appropriate volume of this reagent (add protease inhibitor within a few minutes before use) in the culture plate and bottle for 3-5 min. During this period, the culture plate and bottle were shaken repeatedly to make the reagents fully contact the cells.
  • C. Scrape the cells and reagents with a cell scraper and collect them in a 1.5 mL centrifuge tube.


2. For suspension cells:

Collect the cells by centrifugation, add about 250 uL of this reagent (add protease inhibitor within a few minutes before use) per 6-well plate cells, and shake.

Ice bath for 30 minutes, during which time, pipette with a 200uL pipette several times every 10 minutes to ensure that the cells are completely lysed.
 

  1. Centrifuge at 12000g for 5 min, and collect the supernatant, which is the total protein.


Precautions:
For tissue samples:
Approximately 1mL of this reagent is added to lyse every 50mg tissue. For tissues such as cartilage and skin, the dosage of reagents can be appropriately reduced to increase the protein concentration.

For cultured cell samples:

1. Under normal cell density, add about 250uL of lysate to each 6-well plate cell.
 

  1. It may appear thicker during cracking. It can be pipetted repeatedly until it is liquid. If it has been thicker, you can use a vortex to shake or add an appropriate amount of lysate.
  2. This reagent is harmful to the human body, please protect it appropriately.
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