產(chǎn)品詳情

  • 產(chǎn)品名稱：EZLys(TM)BacterialProteinExtractionReagent

  • 產(chǎn)品型號：
  • 產(chǎn)品廠商：Biovision
  • 產(chǎn)品文檔：
你添加了1件商品 查看購物車
簡單介紹：
EZLys(TM)BacterialProteinExtractionReagent
詳情介紹：
Brand EZLys?
Characteristics A proprietary formulation prepared in a 50 mM Tris pH8.0 buffer and designed for gentle disruption of cell membrane to extract proteins. It provides a simple, rapid, low cost method for releasing expressed target protein for preparation, purification, or other applications. The EZLystm reagent is an optically transparent, non-denaturing formulation which can be easily removed by dialysis. It is also useful for the preparation of high purity inclusion bodies for subsequent purification. This reagent does not interfere with typical protein immobilization or cross linking and is suitable for use in the BCA or Bradford protein assays. Ideal pH range is pH 5 -10. The 100 mL and 500 mL reagent are sufficient for protein extraction from 25 g and 125 g of cell paste.
Application Notes Note: Pellet contains inclusion bodies. If desired, save the pellet for inclusion body purification.)
For Inclusion Body Fraction
1) Resuspend the pellet (from Step A4) in the same volume of EZLys reagent which was used to resuspend the original cell pellet by pipetting up and down and vortexing to obtain an even suspension. Must completely resuspend the pellet.
2) Add lysozyme to a final concentration of 1 KU/mL, mix gently and incubate at room temperature for 10 min.
3) Centrifuge at 5000g x 15 min. at room temperature. Carefully remove supernatant.
4) Resuspend the pellet with 1:10 diluted EZLys using about 8 mL per gram of original wet cell pellet and mix by vortex, centrifuge at 5000g x 15 min. at room temperature.
5) Repeat the step 4) above two more times.
6) Take aliquot for SDS-PAGE. Pellet the inclusion bodies at 16000g x 15 min at 4 °C. Remove supernatant. The inclusion bodies can be stored at -80 °C or use for further applications.
Protocol Note: Pellet contains inclusion bodies. If desired, save the pellet for inclusion body purification.) B. For Inclusion Body Fraction 1) Resuspend the pellet (from Step A4) in the same volume of EZLys reagent which was used to resuspend the original cell pellet (Step A2) by pipetting up and down and vortexing to obtain an even suspension. Must completely resuspend the pellet. 2) Add lysozyme to a final concentration of 1 KU/mL, mix gently and incubate at room temperature for 10 min. 3) Centrifuge at 5000g x 15 min. at room temperature. Carefully remove supernatant. 4) Resuspend the pellet with 1:10 diluted EZLys using about 8 mL per gram of original wet cell pellet and mix by vortex, centrifuge at 5000g x 15 min. at room temperature. 5) Repeat the step 4) above two more times. 6) Take aliquot for SDS-PAGE. Pellet the inclusion bodies at 16000g x 15 min at 4 °C. Remove supernatant. The inclusion bodies can be stored at -80 °C or use for further applications.
Restrictions For Research Use only
Format Liquid
Storage 4 °C
Expiry Date 12 months