Protein Extraction Reagent (Tris) (5X)

Details for Product No. ABIN2344806, Supplier: Log in to see
Reactivity
Bacteria
Application
Immunoassay (IA)
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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 • Available in a phosphate buffer formulation (AKR-181) 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.
Supplier Images
Purification (Purif) image for Protein Extraction Reagent (Tris) (5X) (ABIN2344806) Comparison of Sonication with Bacterial Protein Extraction Reagent. Pelleted His-RFP ...