263454 Detergent-OUT™, SDS Removal Kit

263454
  
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      Overview

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      This product has been discontinued.





      A simple and rapid column based method to remove free SDS from protein solutions. Simply load protein solutions onto column and spin. The detergent is retained by the column matrix and the protein is collected in a small volume. Recovery of protein is generally ≥95%, though some highly hydrophobic proteins may be retained by the column. An SDS test kit is provided for determining detergent removal efficiency. Offered as a mini kit with the capacity to remove 2 mg of SDS from solution or as a medi kit with the capacity to remove up to 10 mg of SDS from the protein solution.
      Catalogue Number263454
      Brand Family Calbiochem®
      Materials Required but Not Delivered Chloroform
      Centrifuge
      15 ml centrifuge tubes
      References
      Product Information
      Form10 Columns
      FormatColumn
      Kit containsDetergent-OUT™ SDS-300 column, SDS Detection Reagents, and a user protocol.
      Applications
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Intended useThe Calbiochem® Detergent-OUT™, SDS Removal Kit is a simple and high performance method for removing free SDS from protein solutions. It offers advantages over equilibrium dialysis, electro-dialysis, and ion exchange methods. This method removes free SDS without substantial loss or dilution of the protein solution.
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Storage ConditionsUpon arrival store the entire contents of the kit at 4°C.
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Kit containsDetergent-OUT™ SDS-300 column, SDS Detection Reagents, and a user protocol.
      Specifications

      Documentation

      Detergent-OUT™, SDS Removal Kit Certificates of Analysis

      TitleLot Number
      263454
      User Protocol

      Revision14-August-2007 JSW
      Form10 Columns
      FormatColumn
      StorageUpon arrival store the entire contents of the kit at 4°C.
      Intended useThe Calbiochem® Detergent-OUT™, SDS Removal Kit is a simple and high performance method for removing free SDS from protein solutions. It offers advantages over equilibrium dialysis, electro-dialysis, and ion exchange methods. This method removes free SDS without substantial loss or dilution of the protein solution.
      Principles of the assayTo remove SDS from protein preparations the protein solution is loaded onto the Detergent-OUT™ Spin column and centrifuged to collect the sample. SDS that is tightly bound to the protein is recovered with the protein solution. Using this method it is possible to achieve a molar ratio of 1:1.1 (protein:SDS). Each Detergent-OUT™ SDS-300 Column will remove up to 10 mg SDS, provided that both the protein solution and the column are prepared in water. The kit is also supplied with SDS detection reagents for determining efficiency of SDS removal.
      Materials provided• Detergent-OUT™ SDS-300 Column (Kit Component No. KP13605): 10 columns
      • Blue Dye (Kit Component No. KP13606): 1 bottle, 30 ml
      • Dye Extraction Buffer (Kit Component No. KP13607): 1 bottle, 15 ml
      Materials Required but not provided Chloroform
      Centrifuge
      15 ml centrifuge tubes
      Precautions and recommendations• Optimal results are obtained when the Detergent-OUT™ SDS-300 Columns and protein solution are prepared in water or phosphate buffers. The Detergent-OUT™ SDS-300 Columns may also be equilibrated with other types of buffers. DO NOT equilibrate the columns with organic- or Tris-based buffers or borate buffers.
      Detailed protocolProtocol for SDS Removal

      1. Invert the Detergent-OUT™ SDS-300 Column several times to resuspend the resin material. Remove the tip of the column and allow the liquid to drain.

      2. Equilibrate a Detergent-OUT™ SDS-300 Columns by adding 1 ml of water (preferred) or other appropriate buffer to the column. Allow the buffer drain into a collection tube and discard. Repeat 3 times.

      3. Place the equilibrated Detergent-OUT™ SDS-300 Column in a 15 ml centrifuge tube and centrifuge at 1000 x g for 20-30 s. Discard the liquid collected in the centrifuge tube.

      4. Place the Detergent-OUT™ SDS-300 Column back in the 15 centrifuge tube and carefully apply the protein solution to the center of the column; incubate for at room temperature for 5 min.

      5. Centrifuge the tube containg the Detergent-OUT™ SDS-300 Column at 1000 x g for 20-30 s. Collect the protein solution from the tube. The protein solution is substantially free of SDS.

      Protocol for Determining Efficiency of SDS Removal

      1. Mix 1-5 µl of the SDS-free protein solution from above with 2 ml Blue Dye and 1 ml Dye Extraction Buffer. Add 2 ml chloroform to the mix and invert the tube 5-6 times to mix. Prepare a control containing 1-5 µl water; treat the water control in the same manner as the protein solution.

      2. Incubate the mixture at room temperature for 5 min. If the chloroform layer extracts color, it is indicative of the presence of SDS in the sample.

      3. Measure the absorbance of the chloroform layer at 600 nm. Do not use disposable cuvettes. Use only quartz or glass cuvettes. By comparing the absorbance of the chloroform layer of the control, you can determine the efficiency of the SDS removal. Up to a two-fold increase in the absorbance over the control (containing no SDS) may simply represent elution of SDS tightly bound to the protein. If the absorbance is substantially increased (beyond two-fold), it is an indication of the elution of free SDS. By preparing calibration plots for protein and SDS concentration, the protein:SDS molar ratio can be determined.
      Registered TrademarksCalbiochem® is a registered trademark of EMD Biosciences, Inc.
      Interactive Pathways™ is a trademark of EMD Biosciences, Inc.
      Detergent-OUT™ is a trademark of Geno Technology, Inc.