For the rapid isolation of inclusion body proteins

  • All-in-one solution for inclusion body protein isolation and purification
  • Complete kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
  • Isolate up to 12 mg of protein per spin column
  • Fast and convenient spin column protocol

ItemsCat. #Size
ProteoSpin™ Inclusion Body Isolation Maxi Kit 17700 4 preps

ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit

This kit provides everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting.

The procedure is efficient and streamlined and can process up to 12 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.

About Inclusion Bodies

Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are tyically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these sub-cellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time consuming and not cost effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography - all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.

Kit Specifications
Maximum Culture Volume
100 mL
Yield from 100 mL of Culture
Up to 4 mg
Minimum Elution Volume
4 mL
Time to Process 1 Sample
1 hour

Storage Conditions
The Cell Lysis Reagent and the IB Solubilization Reagent should be stored at 4°C upon receipt of the kit. All other unopened solutions should be stored at room temperature. Once opened, the solutions should be stored at 4°C when not in use, except for the Basic and Acidic Binding Buffers, which should be stored at room temperature.


Kit Components
Component Cat. 17700 (4 preps)
Wash Solution C 130 mL
Wash Solution N 130 mL
Binding Buffer A 20 mL
Binding Buffer N 2 x 30 mL
Protein Neutralizer 4 mL
Cell Lysis Reagent 110 mL
IB Solubilization Reagent 50 mL
Syringes (10 mL, Luer-Lok™ Tip) 4
Needles (18G x 1½ inch) 4
Maxi Spin Columns (filled with SiC)
inserted into 50 mL collection tubes
4
Final Elution Tubes, 50 mL 4
Product Insert 1

Title CP40 from Corynebacterium pseudotuberculosis is an endo-β-N-acetylglucosaminidase
Journal BMC Microbiology. 2016.
Authors Shadnezhad A, Naegeli A, Collin M.
Title Proteome of Human Calcium Kidney Stones.
Journal Urology. 2010.
Authors Canales BK, Anderson L, Higgins L, Ensrud-Bowlin K, Roberts KP, Wu B, Kim IW, Monga M.