219468 CB-Protein Assay™ Kit

219468
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      Overview

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      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      219468-1KIT
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          Glass bottle 1 kit
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          Description
          OverviewA simple and rapid method for the estimation of protein concentration. Based on an improved Coomassie dye reagent. Reaches a stable endpoint in 5 minutes. Compatible with reducing reagents, but not suitable for use with solutions containing detergents.
          Catalogue Number219468
          Brand Family Calbiochem®
          Application Data
          Materials Required but Not Delivered Disposable polystyrene cuvettes
          Assay tubes
          References
          Product Information
          Form500 tube assays or 5000 microwell plate assays
          Formatcuvette or 96-well plate
          Kit containsCB-Protein Dye, Albumin Standard, and a user protocol.
          Applications
          Biological Information
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          R PhraseR: 36/37/38

          Irritating to eyes, respiratory system and skin.
          S PhraseS: 26-36

          In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
          Wear suitable protective clothing.
          Product Usage Statements
          Storage and Shipping Information
          Ship Code Blue Ice Only
          Toxicity Multiple Toxicity Values, refer to MSDS
          Storage +2°C to +8°C
          Storage ConditionsUpon arrival store entire contents of the kit at 4°C.
          Do not freeze Ok to freeze
          Packaging Information
          Transport Information
          Supplemental Information
          Kit containsCB-Protein Dye, Albumin Standard, and a user protocol.
          Specifications

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          219468
          User Protocol

          Revision21-July-2010 RFH
          Form500 tube assays or 5000 microwell plate assays
          Formatcuvette or 96-well plate
          StorageUpon arrival store entire contents of the kit at 4°C.
          Principles of the assayThe CB-Protein Assay™ Kit offers an improvement over the well-known Bradford Coomassie dye-binding assay. This assay is based on a single Coomassie dye reagent. In an acidic medium, protein binds with Coomassie dye and produces a blue protein-dye complex that can be measured at 595 nm. The test can be performed using as little as 1 µl protein per sample.

          Protein + Coomassie in acidic medium Blue Color Protein-Dye Complex

          Standard Curve for the CB-Protein Assay™ Kit shows a substantially linear relationship between absorbance by density of protein-dye complex and the protein concentration.
          Materials provided• CB-Protein Assay™ Reagent (Kit Component No. KP15201): 1 x 500 ml
          • Bovine Serum Albumin Standard (2 mg/ml) (Kit Component No. KP15202): 1 x 5 ml
          Materials Required but not provided Disposable polystyrene cuvettes
          Assay tubes
          Precautions and recommendations Allow the CB-Protein Assay™ Reagent to warm to room temperature. Prior to use, mix the reagent gently by inverting the bottle several times. DO NOT SHAKE THE BOTTLE.
          Staining of glassware: clean glassware with methanol.
          Effect of temperature: consistent results are obtained when the CB-Protein Assay™ Kit is at room temperature. Allow components to warm to room temperature prior to use.
          Preparation• Preparation of diluted protein standard: For minimizing interference, it is important to prepare the appropriate diluted protein standard in the same diluent used for the test protein sample.

          Standard Protocol (2000-25 µg protein/ml)

          Table 1: Standard Protocol



          Micro Protocol
          Prepare a 100 µg protein/ml stock solution by mixing 25 µl (2 mg/ml stock) albumin solution and 475 µl diluent. Use this stock for preparing diluted protein standard for the micro protocol assay. For minimizing interference, it is important to prepare the standard in the same diluent used for the test protein sample.

          Table 2: Micro Protocol

          Detailed protocol
          Protein assay can be performed either by the standard protocol in the concentration range 100-1,000 µg protein/ml BSA or by micro-assay protocol for dilute protein solution in the working range of 1-25 µg/ml.

          Standard - Test Tube Assay: (Protein Concentration Range 100-1000 µg/ml)

          Perform assay with duplicate protein standards and samples. Read Precautions Recommendations.
          1. Transfer 0.05 ml of diluted standards and test samples in the assay tubes. Set up a blank containing 0.05 ml of protein solution diluent.
          2. Add 1 ml of CB-Protein Assay™ reagent into each tube and mix well. Allow the tubes to incubate at room temperature for at least 5 min, but no more than 1 h.
          3. Read absorbance of the assay tubes at 595 nm.
          4. Subtract reading of the blank with the test samples and plot a standard curve for determination of the protein concentration of unknown samples.

          Standard - 96-Well Plate Assay: (Protein Concentration Range 100-1000 µg/ml)

          Perform assay with duplicate protein standards and samples. Read "Preparation Before Use".
          1. Transfer 10 µl of diluted standards and test samples into plate wells. Set up a blank containing 10 µl of protein solution diluent.
          2. Add 200 µl of CB-Protein Assay™ reagent into each well and mix well. Allow the wells to incubate at room temperature for at least 5 min, but no more than 1 h.
          3. Read absorbance of the assay tubes at 595 nm.
          4. Subtract reading of the blank with the test samples and plot a standard curve for determination of the protein concentration of unknown samples.
          Micro Protocol: Test Tube Assay (Protein Conentration Range 1-25 µg/ml)

          Perform assay with duplicate protein standards and samples. Read Precautions Recommendations.
          1. Transfer 0.5 ml of diluted standards and test samples in the assay tubes. Set up a blank containing 0.5 ml of protein solution diluent.
          2. Add 0.5 ml of CB-Protein Assay™ reagent into each tube and mix well. Allow the tubes to incubate at room temperature for at least 5 min, but no more than 1 h.
          3. Read absorbacne of the assay tubes at 595 nm.
          4. Subtract reading of the blank with the test samples and plot a standard curve for determination of the protein concentration of unknown samples.

          Micro Protocol: Plate Assay (Protein Concentration Range 1-25 µg/ml)

          Perform assay with duplicate protein standards and samples. Read Precautions Recommendations.
          1. Transfer 100 µl of diluted standards and test samples into plate wells. Set up a blank containing 100 µl of protein solution diluent.
          2. Add 100 µl of CB-Protein Assay™ reagent into each well and mix well. Allow the wells to incubate at room temperature for at least 5 min, but no more than 1 h.
          3. Read absorbance of the assay tubes at 595 nm.
          4. Subtract reading of the blank with the test samples and plot a standard curve for determination of the protein concentration of unknown samples.
          Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
          Triton® is a registered trademark of Dow Chemical Company
          Tween® and Brij® are registered trademarks of ICI Americas, Inc.
          Interactive Pathways™ is a trademark of EMD Chemicals, Inc.
          CB-Protein Assay™ Kit is a trademark of Geno Technology, Inc.