Catalog

10100

ProteoSpin CBED Micro Kit

( 25 preps )
Norgen Biotek

479.99$

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Additional description

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

Product Description

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 technology. 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 employsa proprietarymatrix, whichacts as an ion exchanger and is able to concentrate proteinsmany fold as well as remove salts. Briefly, pH Binding Buffer is added to the protein sample and the sample is applied toNorgen's column (BIND). Under these conditions the proteins will bind to the column while salts and other contaminants are removed inthe flowthrough. The bound proteins are then washed to remove any remaining impurities(WASH). Lastly, theclean proteins are elutedinto a small volume ofthe provided Elution Buffer or intoother optional elutionbuffers(user-provided)(ELUTE).Please see theprocedure flowchart to theright.
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.TheProteoSpinTM CBED Micro Kittherefore 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 ofeither 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.

Specifications

The concentrated proteins are ready for any downstream proteomic applications including:

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.

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 48µg of the input protein was recovered, corresponding to a 38X protein concentration based on mass.

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, CaCl2or 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 expecteddue 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.

APPLICATIONS

Kit Specifications
Maximum Protein Input
50 µg
Protein Recovery*
Up to 99%
Minimum Elution Volume
30 µL
% Salt Removal
Up to 99%
Volume Concentration Factor
Up to 100X
Time to Process 10 Samples
20 minutes
*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.