Catalog

10200

ProteoSpin Detergent Clean-Up Micro Kit

( 25 preps )
Norgen Biotek

479.99$

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Additional description

ProteoSpin Detergent Clean-Up Micro Kit (Cat# 10200 )

For the rapid and simple removal of detergents from protein samples

  • Concentrate protein samples and remove detergents at the same time
  • Effectively remove a wide range of detergents
  • Fast processing time, rapid spin-column format

Product Description

The ProteoSpin™ Detergent Clean-Up Micro Kit removes detergents from protein samples while concentraing the sample at the same time. The kit employs anovel spin column technology that offers convenience, speed, and ease-of-use. The kit can remove trace amounts of detergents from protein samples while maintaining high protein recovery.All types of detergents including ionic, non-ionic and zwitterionic detergents can be effeciently removed in a very short time, eliminating the need for for labourious procedures such as dialysis.The kit is designed to remove detergents in protein solutions either in their free form or bound form, as when complexed with the protein. Also,unlike other commercially available kits that can onlyremove a singledetergent andoften do notremove denaturants, the ProteoSpin™ Detergent Clean-Up Micro Kit removes a variety of detergents and denturants that may be used during the purification of the protein sample while providing high protein recovery.

The kit employs a prorietary matrix to remove detergents from protein samples. 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 whiledetergents 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.

No furtherpurificationof the protein sample is required after detergent removal, and it can be used directly in a variety of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, and NMR spectroscopy. This kit also offers the additional advantage of fractionating basic proteins from a protein solution, thereby allowing foradditional purification of that basic proteins from other contaminating acidic proteins.

FEATURES AND BENEFITS

  • Remove a variety of detergents - Detergents including SDS, Triton® X-100, CHAPS, NP-40, Triton® X-114 and Tween 20 can be removed using the kit.
  • Effective detergent removal - Detergent removal of greater than 95% for many protein samples, allowing for detergent-sensitive tryptic digestion. Can also remove detergents that are complexed with proteins.
  • Complete kit - The ProteoSpin™ Total Protein Detergent Clean-Up Micro Kit contains all the solutions and columns for the processing of 25 total protein samples. Each spin column is able to process 50 µg of either acidic or basic protein
  • Fast and easy processing - Preparation time for 12 samples is only 20 minutes.
  • Eluted proteins can be used directly in downstream applications - No need for additional sample preparation procedures for downstream applications. Concentrate protein sample and remove detergents at the same time.
  • No detergent carryover - Each sample is processed individually using its own spin column, resulting in no carryover or column bleed from sample to sample.
  • Ready-to-use columns - No purging, swelling of resin, or lengthy activation steps required.

Specifications

The purified proteins are suitable for a number of downstream applications such as:

  • 2D SDS-PAGE
  • Whole protein mass spectrometry
  • Isoelectric focusing
  • X-ray crystallography
  • NMR spectroscopy
  • Protein microarrays

Table 1. Efficient Removal of SDS

Removal of SDS with the ProteoSpinTM Detergent Clean-Up Micro Kit is greater than 99%. To evaluate this, 50 µg samples of BSAwere spiked with 0.1% SDSand processed using the kit. The protocol was followed, and the proteins were bound, washed and eluted as described. The purified protein was then assayed for SDS using the Detergent-OUT kit by Geno Technologies, Inc, and SDSremoval was found to be greater than 99%.

Figure 1. High Protein Recovery in the Presence of Detergents. To investigate the recovery of proteins in the presence of different amounts of detergent, 50 µg samples of BSA were prepared insolution and spiked with increasing amounts of SDS (0%–1.0%). The protein samples were then purified using the ProteoSpinTM Detergent Clean-Up Micro Kit according to the bind, wash and elution procedure to remove the SDS. The percentage of recovery of BSA was then determined using the Bradford Assay. High protein recovery was obtained evenwhen the original SDS concentration was as high as 1%.

APPLICATIONS

Kit Specifications
Maximum Protein Input
50 µg
Protein Recovery
80 - 95%
Column Binding Capacity
50 µg
% Detergent Removal
> 95%
Minimum Elution Volume
30 µL
Time to Process 12 Samples
20 minutes


ProteoSpin Detergent Clean-Up 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.