Catalog

10300

ProteoSpin Inclusion Body Isolation Micro Kit

( 20 preps )
Norgen Biotek

498.09$

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Additional description

ProteoSpin Inclusion Body Protein Isolation Micro Kit (Cat# 10300 )

For the rapid isolation of inclusion body proteins - cells to gels in 60 minutes

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

Product Description

The ProteospinTM Inclusion Body Isolation Micro Kit is designed for the purification of recombinant proteins expressed in bacteria as inclusion bodies.Bacteria are widely used for the expression of different recombinant protein. However, 70-80% of the proteins expressed in E. coli by recombinant techniques are contained in inclusion bodies (i.e., protein aggregates). Procedures for the purification of the expressed proteins from inclusion bodiesare often labour-intensive, time consuming and not cost effective.Norgen's ProteospinTM Inclusion Body Isolation Micro Kit offers many advantages over these other procedures. The kit provides the essential reagents for cell disruption, inclusion body solubilisation and purification using spin column chromatography -all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.

The ProteospinTM Inclusion Body Isolation Micro Kit uses a proprietary Cell Lysis Reagent to selectively lyse the cells and release inclusion bodies in their solid form. Next,inclusion bodies are dissolved and their contents released using the IB Solubilization Reagent. Inclusion body proteins are then further purified by loading onto spin columns containing Norgen's proprietary resin (BIND). Non-specifically bound components in the solution can then be washed from the column, leaving the inclusion body protein bound to the resin (WASH). These specific proteins can then be eluted into 50 µL of the supplied Elution Buffer (ELUTE).Please see the procedure flowchart to the right.

The process is efficient and streamlined.The kit can process up to 12 samples in only 60 minutes.Each spin column is able to recover up to 50 µg of acidic or basic protein. This kit is often used to screen recombinants prior to scale up. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, mass spectrometry analysis, and other applications

FEATURES AND BENEFITS

  • Three kits in one for acidic and basic proteins - The kit provides essential reagents for cell disruption, inclusion body solubilization and purification using spin-column chromatography
  • Gentle disruption procedure - Cell lysis is accomplished through non-ionic detergent chemical disruption. Passing cells through a needle eliminates the need for cell sonication or other forms of mechanical disruption, producing proteins that are >95% pure.
  • Long shelf life - Enzyme-free process with stable solutions for longer shelf life.
  • Fast and easy processing- Process up to 12 samples in only 60 minutes using a rapid spin-column procedure
  • Ready-to-use columns - No column preparations required.
  • Proteins suitable for downstream applications- The purified proteins are suitable for a number of downstream applications such as SDS-PAGE, 2D gels, mass spectrometry analysis or other procedures

Specifications

Purified proteins can be used in a number of downstream applications including:

  • SDS-PAGE analysis
  • Western blot
  • ELISA
  • Refolding experiments
  • Mass spectrometry
  • Further purification and scale up

Figure 1. Isolation of Acidic Recombinant Protein. Small (1.5 mL) cultures of BL21 (DE3) pLysS bacteria expressing a 50 kDa chimeric protien (acidic) were pelleted. Cells were lysed and inclusion bodies separated and dissolved using theProteoSpin™ Inclusion Body Protein Isolation Micro Kit.The resulting protein was bound to the ProteoSpinTM column, washed and eluted in 50 µL of the provided elution buffer. The eluted protein samples were analyzed in 12.5% polyacrylamide gels, which were run for 45 minutes at 200 V/6.5 cm. The protein bands were made visible by staining with Coomassie Blue R-250.

Figure 2. Isolation of Basic Recombinant Protein (RNase A). Small (1.5 mL) cultures of BL21 (DE3) pLysS bacteria expressingRNaseA (basic) were pelleted. Cells were lysed and inclusion bodies separated and dissolved using theProteoSpin™ Inclusion Body Protein Isolation Micro Kit.The resulting protein was bound to the ProteoSpinTM column, washed and eluted in 50 µL of the provided elution buffer. The eluted protein samples were analyzed in 12.5% polyacrylamide gels, which were run for 45 minutes at 200 V/6.5 cm. The protein bands were made visible by staining with Coomassie Blue R-250.

Figure 3. Efficient Isolation of Inclusion Body Proteins Using ProteoSpin Inclusion Body Protein Isolation Kits. Following gene expression, 1.5 mL cultures of BL21 (DE3) pLysS bacteria expressingproteins of various molecular weights were pelleted by centrifugation. Cells were lysed and inclusion bodies separated and dissolved using theProteoSpin™ Inclusion Body Protein Isolation Micro Kit.The resulting proteins were bound to the ProteoSpinTM columns, washed and eluted in 50 µL of the provided elution buffer. The eluted protein samples were analyzed in 12.5% polyacrylamide gels, which were run for 45 minutes at 200 V/6.5 cm. The protein bands were made visible by staining with Coomassie Blue R-250. Lane labels indicate the size of each protein in kDa.

APPLICATIONS

Kit Specifications
Maximum Culture Volume
1.5 mL
Yield from 1.5 mL
Up to 50 µg
Minimum Elution Volume
30 µL
Time to Process 12 Samples
60 minutes
ProteoSpin Inclusion Body Protein Isolation Micro Kit Contents
1. Cell Lysis Reagent
2. Inclusion Body Solubilization Reagent
3. Column Activation and Wash Buffer for Acidic Proteins
4. pH Binding Buffer for Acidic Proteins
5. Column Activation and Wash Buffer for Basic Proteins
6. pH Binding Buffer for Basic Proteins
7. Elution Buffer
8. Neutralizer
9.Micro Spin Columns (20)
10. Collection Tubes (20)
11. Elution Tubes (20)
12. Syringes (20)
13. Needles (20)
14. Product Insert


Storage Conditions
The Cell Lysis Reagent and the IB Solubilization Reagent should be stored at 4°Cupon 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.