產(chǎn)品詳情

  • 產(chǎn)品名稱：ProteinExtractionReagent(Phosphate)(5X)

  • 產(chǎn)品型號：
  • 產(chǎn)品廠商：CellBiolabs
  • 產(chǎn)品文檔：
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ProteinExtractionReagent(Phosphate)(5X)
詳情介紹：
Characteristics The Bacterial Protein Extraction Reagent contains a gentle, nonionic detergent formulation which quickly extracts functional, recombinant protein from E. coli without mechanical disruption. Other benefits include the following: ? Designed specifically for mild extraction of functional, recombinant proteins from E. coli without mechanical disruption ? Produces high yields in a single-step, 15-minute protein extraction (entire procedure takes less than 30 minutes) ? Designed for small-scale or large-scale protein extraction ? Protease inhibitors, reducing agents, Lysozyme and/or DNase I are easily added to improve lysis/recovery ? Compatible with 6xHis and GST affinity purification systems ? Phosphate buffer formulation for direct labeling and conjugation ? Recovered lysates are compatible with SDS-PAGE, BCA and Bradford Protein Assays ? Can be used with frozen or recently prepared E. coli pellets
Background Bacteria are widely used for recombinant protein expression. However, mechanical cell disruption methods (e.g. sonication, steel beads, high-shear) are often time-consuming or limited by sample volume, sample number, disruption efficiency, protein functionality, and total yield.
Application Notes Optimal working dilution should be determined by the investigator.
Comment

  • Created for small-scale or large-scale protein extraction
  • Compatible with 6xHis and GST affinity purification systems
  • Designed specifically for mild extraction of functional, recombinant proteins from E. coli without mechanical disruption

Assay Procedure
  1. Pellet bacterial cells by centrifugation at 5000 x g for 10?minutes.
  2. Aspirate the media from the pellet.
  3. Resuspend the cell pellet with 1X Bacterial Protein Extraction Reagent (see Preparation of Reagent Section). Add 4?mL per gram of cell pellet. Triturate cells up and down until the solution is homogeneous.
  4. Incubate 15?minutes at room temperature.
  5. Centrifuge the lysate 12000 x g for 10?minutes.
  6. Carefully transfer the supernatant to another tube, making sure not to disturb the insoluble pellet. Note: If desired recombinant protein is not soluble (forming inclusion bodies), adjust the expression conditions or use an inclusion body solubilization reagent (such as #AKR-110 for GST fusion proteins).
Restrictions For Research Use only
Format Liquid
Concentration 5 X
Storage RT
Storage Comment Store at room temperature for up to 1 year from date of receipt.