For high-yield extraction of soluble proteins from insect cells.

Thermo Scientific I-PER Insect Cell Protein Extraction Reagent enables gentle and effective extraction of soluble protein from baculovirus-infected insect cells grown in suspension or monolayer culture. The baculovirus insect cell expression system is an efficient and popular system for production of recombinant (eukaryotic) proteins in cell culture. Proteins expressed in baculoviral systems can be used for structural analyses, biochemical assays and a variety of other applications. I-PER Reagent maintains functionality of extracted proteins and is directly compatible with downstream applications such as protein assays, Western blotting and 6xHis-tagged protein purification.

Highlights:

• Gentle extraction – optimized, mild non-ionic detergent provides maximum extraction of soluble proteins from insect cells
• Effective – provides better protein extraction than sonication
• Compatible – downstream compatibility with Western blotting, 6xHis-tagged protein purification, protein assays and ion-exchange chromatography
• Flexible – useful for protein extraction from suspended or adherent cultured insect cells

Efficient extraction of recombinant proteins from insect cells. Cell lysates were prepared from infected Sf9 cells using Thermo Scientific I-PER Insect Protein Extraction Reagent. Normalized amounts of total, insoluble and soluble protein were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) before Western blot analysis.

Affinity purification of 6xHis Cyclin B1 from insect cell extract. Baculovirus-infected Sf9 cells were harvested and lysed with Thermo Scientific I-PER Reagent. The extract was directly loaded onto a nickel-chelated agarose (IMAC) column and purified. Protein samples were separated by SDS-PAGE, and the gel was stained with GelCode Blue Stain Reagent (Part No. 24590).

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