Catalog

17000

ProteoSpin CBED Maxi Kit

( 4 preps )
Norgen Biotek

235.47$

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Additional description

CBED (Concentration, Buffer Exchange and Desalting) Maxi Kit (Cat# 17000 )

For rapid and efficient concentration and desalting of protein samples

  • Protein recovery up to 99%
  • Up to 99% removal of salts
  • Fast processing time, rapid spin-column format

Product Description

The ProteoSpinTM CBED Maxi Kit provides a fast and a simple procedure for concentration, buffer exchange and desalting of protein samples based on spin column chromatography. The ProteoSpin™ CBED Maxi 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 described.

The kit employs an innovative separation matrix, whichacts as an ion exchanger, and is able to concentrate proteinsmany fold as well as removes 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.
The kit can efficiently process 4 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 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 -Process 4 samples in only 20 minutes, compared to other lengthy concentrating and desalting methods.
  • Efficient salt removal - Remove up to 99% of salts
  • High protein recovery - Recover up to 99% of protein input
  • 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 Maxi Kit

Input Volume
Output Volume
Concentration Factor
20 mL
4 mL
5X
60 mL
4 mL
15X
100 mL
4 mL
25X

To demonstrate the ability of the ProteoSpinTM CBED Maxi Kit to concentrate protein samples, 8 mg of BSA was spiked into 20,60 and100 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 4 mL of the provided Elution Buffer. The20 mL input was therefore concentrated 5X, the60 mL input was concentrated 15X, and the100 mL input was concentrated 25X.

Table 2: Examples of BSA Mass Concentration using the ProteoSpin™ CBED Maxi Kit

Input Concentration
Output Concentration
Concentration Factor
8 mg / 20 mL
7.89 mg / 4 mL
5X
(0.4 mg/mL)
(1.97 mg/mL)

To further demonstrate the ability of the ProteoSpinTM CBED Maxi Kit to concentrate protein samples, 8 mg of BSA was spiked into20 mL of a 50 mM sodium acetate solution (pH 4.5). The sample was processed as per the provided protocol, and was eluted into 4 mL of the provided Elution Buffer. The amount of recovered protein was then determined using the BioRad Protein Assay. It was found that 7.89 mg of the input protein was recovered in 4 mL, corresponding to a 5X protein concentration based on mass.

Figure 1. High Protein Recovery. Increasing amounts of BSA (1, 2, 4, 6 and 8 mg) were spiked into 20 mLof a 50 mM sodium acetate solution, and were concentrated using the ProteoSpinTMCBED Maxi Kit. The provided protocol was followed, and the concentrated BSAwas eluted into 2 separate 4 mLelutions using the provided Elution Buffer. The elutions were then combined, and the amount of protein recovered in each elution was determined using the BioRad ProteinAssay.The ProteoSpin™ CBED Maxi Kit allows for high proteinrecoveries of up to 90%.

APPLICATIONS

Kit Specifications
Maximum Protein Input
8 mg
Minimum Protein Input
0.25 mg
Protein Recovery*
Up to 99%
Column Binding Capacity
8 mg
% Salt Removal
Up to 99%
Volume Concentration Factor
Up to 25X
Minimum Elution Volume
4 mL
Time to Process 4 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 Maxi 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. Maxi Spin Columns (4)
8. Elution Tubes (4)
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.