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Norgen Biotek Corp. is dedicated to providing our customers with first class sample preparation kits for RNA, microRNA, DNA and protein purification, clean-up and concentration and to providing dedicated and expert support services to our customers and partners worldwide.
As a testament of our commitment to quality, Norgen is proud to be an ISO 9001:2008 and ISO 13485:2003 registered company.
It is a pleasure to serve you and we look forward to working closely with you to provide the best sample preparation products for your specific needs. |
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Please contact us at:
info@norgenbiotek.com
Norgen Biotek Corp.
3430 Schmon Parkway,
Thorold
Ontario, Canada, L2V 4Y6
Tel: (905) 227-8848
Fax: (905) 227-1061 |
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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 kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
- Fast and convenient spin column protocol
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The ProteospinTM Inclusion Body Isolation Maxi Kit is a complete kit that 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 for rapid and efficient cell lysis, protein solubilization and on-column protein purification of up to 12 mg of proteins from 100 mL of bacterial culture. No other 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 or extra 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.
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| APPLICATIONS
Purified proteins can be used directly in a number of downstream applications including:
- Mass Spectrometry
- SDS-PAGE analysis
- Refolding Experiments
- Western blot
- ELISA
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Figure 1: No Loss of Proteins when Isolating a Basic Protein. 100 mL of induced bacterial culture expressing a recombinant 30 kD BASIC protein were pelleted and processed using the ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit. Briefly, pelleted cells were lysed, inclusion bodies were separated and subsequently dissolved using the provided Inclusion Body Solubilization 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 culture expressing a recombinant 70 kD ACIDIC protein were pelleted and processed using the ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit. Briefly, pelleted cells were lysed, inclusion bodies were separate and subsequently dissolved using the provided Inclusion Body Solubilization 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 using Norgen'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.
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Kit Specifications
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Maximum Culture Volume
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100 mL
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Yield from 100 mL of Culture
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Up to 12 mg
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Minimum Elution Volume
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4 mL
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Time to Process 1 Sample
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1 hour
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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.
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Order NOW at our online store:
Item |
Cat. No. |
Kit Size |
Buy Online |
ProteoSpin Inclusion Body Isolation Maxi Kit |
17700 |
4 preps |
Buy Now |
Order by Phone: Toll Free (USA & CANADA): 1-866-NORGENB (667-4362) or from anywhere at (905)227-8848.
Order by Email: order by sending an email to orders@norgenbiotek.com.
Order by Fax: Fax your order to (905)227-1061
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