Catalog

17700

ProteoSpin Inclusion Body Isolation Maxi Kit

( 4 preps )
Norgen Biotek

268.07$

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Additional description

ProteoSpin Inclusion Body Protein Isolation Maxi Kit (Cat# 17700 )

For the rapid isolation of inclusion body proteins

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

Product Description

The ProteospinTM Inclusion Body Isolation Maxi Kit is acomplete kitthat 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). This kit is stream- lined forrapid andefficient celllysis, protein solubilization andon-column protein purification of up to 12 mg of proteins from 100 mL ofbacterialculture. Noother kit on the market offers all of these processes in one kit. The purified proteins can be run directly gels or further characterized without any additional steps such as dialysis orextra desalting.

Procedures for the purification of inclusion body proteins are often labour-intensive, time consuming and not cost effective.The ProteospinTM Inclusion Body Isolation Maxi Kit offers many advantages over these other procedures.The 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 tremendous amounts of time and money.

The ProteospinTM Inclusion Body Isolation Maxi 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 6 mL of the supplied Elution Buffer (ELUTE).Please see the procedure flowchart to the right.
The process is efficient and streamlined. Purified recombinant proteins are 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. The kit includes solutions and protocols for use with either acidic or basic proteins.
  • Gentle disruption procedure provides high purity final product - Cell lysis is accomplished through non-ionic detergent chemical disruption. The use of a needle and syringe reduces viscosity and facilitates purification, producing protein often >95% pure. Proteins are then ready for SDS-PAGE, 2D gels, mass spectrometry analysis or other procedures.
  • Convenient process volume - Analyze 100 mL of microbial cell culture yielding 12 mg of protein.
  • Long shelf life - The kit uses an enzyme-free process with stable solutions for longer shelf life.
  • Fast process time - Process time is about 2 hours for a single sample.
  • Ready-to-use columns - No column preparation is required.

Specifications

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

  • Mass Spectrometry
  • SDS-PAGE analysis
  • Refolding Experiments
  • Western blot
  • ELISA

Figure 1: No Loss of Proteins when Isolating a Basic Protein. 100 mL of induced bacterial cultureexpressing a recombinant 30 kDBASICproteinwere pelleted and processed using the ProteoSpin™Inclusion Body Protein Isolation Maxi Kit. Briefly, pelleted cells were lysed, inclusion bodies were separatedand subsequently dissolved using the provided Inclusion BodySolubilization Reagent. Fractions of input, flowthrough, wash and elution were loaded on a 12.5%acrylamide gel. Lane 1 is the input, Lane 2 is the flowthrough from the input, Lane 3 is the wash, Lane 4 is the first elution and Lane 5 is the second elution. As can be seen, proteins are not lost in the flowthrough or wash. Recombinant proteins were efficiently bound to the column and eluted.

Figure 2: No Loss of Proteins when Isolating An Acidic Protein. 100 mL of induced bacterial cultureexpressing a recombinant 70 kDACIDICproteinwere pelleted and processed using the ProteoSpin™Inclusion Body Protein Isolation Maxi Kit. Briefly, pelleted cells were lysed, inclusion bodies were separateand subsequently dissolved using the provided Inclusion BodySolubilization Reagent. Fractions of input, flowthrough, wash and elution were loaded on a 12.5%acrylamide gel. Lane 1 is the input, Lane 2 is the flowthrough from the input, Lane 3 is the wash, Lane 4 is the first elution and Lane 5 is the second elution. As can be seen, proteins are not lost in the flowthrough or wash. Recombinant proteins were efficiently bound to the column and eluted.

Figure 3: Efficient Isolation of Inclusion Body Proteins Using ProteoSpin Inclusion Body Protein Isolation Kits. Following gene expression, inclusion bodies were extracted, solubilized and proteins were purified usingNorgen's Inclusion Body Protein Isolation Kits. Briefly, cells were pelleted and lysed using the provided Cell Lysis Reagent. The inclusion bodies were separated via centrifugation and washing and dissolved using the Inclusion Body Solubilization Reagent. Recovered proteins were analyzed on 12% SDS-PAGE and stained with Coomassie Brilliant Blue R-250. Numbers represent kDa sizes of protein bands. As can be seen, proteins of a wide range of masses are efficiently purified. Lane numbers represent size of protein purified in kDa.

APPLICATIONS

Kit Specifications
Maximum Culture Volume
100 mL
Yield from 100 mL of Culture
Up to 12 mg
Minimum Elution Volume
4 mL
Time to Process 1 Sample
1 hour


ProteoSpin Inclusion Body Protein Isolation Maxi 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. Maxi Spin Columns (4)
10. Collection Tubes (4)
11. Elution Tubes (4)
12. Syringes (4)
13. Needles (4)
14. Product Insert


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.