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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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CBED (Concentration, Buffer Exchange and Desalting) Micro Kit (Cat#
10100 )
For rapid and efficient concentration and desalting of protein samples
- Protein recovery of up to 99%
- Up to 99% removal of salts
- Fast processing time, rapid spin-column format
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The ProteoSpinTM CBED Micro Kit provides a fast and a simple procedure for concentration, buffer exchange and desalting of protein samples based on spin column technoloy. The ProteoSpin™ CBED Micro Kit comes with solutions for concentrating and desalting both acidic and basic proteins. Two procedures, one for acidic proteins and another for basic proteins, are provided.
The kit employs a proprietary matrix, which acts as an ion exchanger and is able to concentrate proteins many fold as well as remove salts. Breifly, pH Binding Buffer is added to the protein sample and the sample is applied to Norgen's column (BIND). Under these conditions the proteins will bind to the column while salts and other contaminants are removed in the flowthrough. The bound proteins are then washed to remove any remaining impurities (WASH). Lastly, the clean proteins are eluted into a small volume of the provided Elution Buffer or into other optional elution buffers (user-provided) (ELUTE). Please see the procedure flowchart to the right.
One major advantage to this kit is there is no molecular weight cut-off, unlike products from other vendors. A broad size range of proteins, from peptides to antibodies can be processed. Salts such as MgCl2, NaCl, KCl, CaCl2, LiCl, and CsCl have poor affinities for Norgen’s proprietary resin and are easily removed from the sample. Another advantage over other vendor products is that the ProteoSpinTM CBED Micro Kit concentrates, removes salts and offers buffer exchange at the same time, whereas other products including dialysis do these processes as separate procedures. The ProteoSpinTM CBED Micro Kit therefore saves considerable time and expense.
Each spin column can process up to 50 µg of protein in up to 5 mL of solution. The kit can efficiently process 12 samples in only 20 minutes using an easy-to use protocol. The percentage of protein recovery is up to 99%, whereas the percentage of salt removal is up to 99%. The kit allows for the concentration of either basic or acidic proteins separately The concentrated proteins are ready for any downstream proteomic applications including 2D SDS-PAGE, Whole protein mass spectrometry, Isoelectric focusing, X-ray crystallography, NMR spectroscopy and Protein microarrays FEATURES AND BENEFITS
- Fast processing time - Efficiently process 12 samples in only 20 minutes compared to other concentrating and desalting methods.
- High yield - Recovery of up to 99% of input protein amount.
- Efficient salt removal - Remove up tp 99% of salts
- No molecular weight cutoff - Based on an ion-exchange mechanism, a broad size range of proteins from peptides to antibodies can be processed.
- Ready-to-use columns - No column preparations required.
- No special storage conditions - All the kit components can be stored at room temperature
- No mixing or formulation - Solutions and protocols are provided for both acidic and basic proteins with no need for formulation.
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| APPLICATIONS
The concentrated proteins are ready for any downstream proteomic applications including:
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Table 1. Examples of BSA Volume Concentration using the ProteoSpin CBED Micro Kit.
To demonstrate the ability of the ProteoSpinTM CBED Micro Kit to concentrate protein samples, 50 µg of BSA was spiked into 1, 3 and 5 mL of a 50 mM sodium acetate solution (pH 4.5). The samples were processed as per the provided protocol, and all samples were eluted into 50 µL of the provided Elution Buffer. The 1 mL input was therefore concentrated 20X, the 3 mL input was concentrated 60X, and the 5 mL input was concentrated 100X.
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Table 2: Examples of BSA Concentration using the ProteoSpin CBED Micro Kit
To further demonstrate the ability of the ProteoSpinTM CBED Micro Kit to concentrate protein samples, 50 µg of BSA was spiked into 2 mL of a 50 mM sodium acetate solution (pH 4.5). The sample was processed as per the provided protocol, and was eluted into 50 µL of the provided Elution Buffer. The amount of recovered protein was then determined using the BioRad Protein Assay. It was found that 48 µg of the input protein was recovered, corresponding to a 38X protein concentration based on mass.
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Table 3. Representative Recovery and Salt Reduction using the ProteoSpin CBED Micro Kit
To investigate salt removal using the ProteoSpinTM CBED Micro Kit, 50 µg samples of BSA were prepared in 1 mL samples of sodium acetate and were spiked with 300 ppm of MgCl2, NaCl, KCl, CaCl2 or LiCl. The salt was removed using the provided protocol, and the proteins were eluted into 2 separate 50 µL elutions of 250 mM ammonium hydroxide. Ammonium hydroxide is not the provided elution buffer, but was required to allow for the analysis of the amount of salt in the elutions. The 2 elutions were combined for analysis. The amount of BSA recovery was determined using the BioRad Protein Assay, and the amounts can be seen in the table above. The BSA recoveries were lower than expected due to the fact that an alternate Elution Solution was used for analysis that was not optimized for recovery. The amount of salt present in the elutions was determined by ICP/MS designed for elemental analysis. As it can be seen, salt removal was up to 99.7% using this kit. |
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Kit Specifications
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Maximum Protein Input
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50 µg
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Protein Recovery*
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Up to 99%
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Minimum Elution Volume
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30 µL
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% Salt Removal
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Up to 99%
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Volume Concentration Factor
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Up to 100X
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Time to Process 10 Samples
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20 minutes
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*Recoveries of up to 99% have been achieved using BSA and the supplied elution buffer. Recovery will vary depending on the protein sample and the elution buffer used.
CBED Micro Kit Contents
1. Column Activation and Wash Buffer for Acidic Proteins
2. pH Binding Buffer for Acidic Proteins
3. Column Activation and Wash Buffer for Basic Proteins
4. pH Binding Buffer for Basic Proteins
5. Elution Buffer
6. Neutralizer
7. Micro Spin Columns (25)
8. Elution Tubes (25)
9. Product Insert
Storage Conditions
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 CBED Micro Kit |
10100 |
25 preps |
Buy Now |
ProteoSpin CBED Micro Kit |
10400 |
50 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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