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152226 AEC Substrate Buffer

This AEC Substrate Buffer is validated for use in Immunoblotting and Immunohistochemistry.

AEC Substrate Buffer MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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152226
  
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      Overview

      Replacement Information
      Description
      OverviewPeroxidase substrate buffer supplied as a 10X concentrate (pH 5.0 when diluted). A diluent for Cat. No. 152224.
      Catalogue Number152226
      Brand Family Calbiochem®
      References
      Product Information
      FormLiquid
      FormulationIn 500 mM acetate buffer, 88 mM H₂O₂, proprietary stabilizers.
      PreservativeNone
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunohistochemistry
      Application NotesImmunoblotting (see comments)
      Immunohistochemistry (see comments)
      Application CommentsStable at 4°C, 18-26°C, and 37°C. When diluted the final pH is ~5.04. Functionality tested by performing immunohistologic stain using the final product.

      Suggested procedure for use of AEC in immunohistochemistry and immunoblotting:

      1. Complete all required incubations with antibodies and HRP-labeled probes, and all necessary wash steps.
      2. Prepare 1X AEC Substrate Buffer: Add 9 parts distilled water to 1 part 10X AEC Substrate Buffer, Cat. No. 152226 (0.5 M acetate buffer containing 88 mM H2O2). Mix well. The final pH of this solution will be ~5.0.
      3. Prepare 1X AEC Substrate Solution: Add 1 part 50X AEC Concentrate (Cat. No. 152224) to 50 parts 1X AEC Substrate Buffer (prepared in Step 2). Mix well.
      4. Following the final wash, completely cover tissue sections with 1X AEC Substrate Solution (prepared in Step 3) and incubate 10-30 min at room temperature. The same conditions apply for immunoblotting. Note: Variables associated with assay conditions will dictate the proper reaction times.
      5. After reaction is complete, wash tissue sections or blotting membranes thoroughly in distilled water.
      6. If tissue sections require a counterstain, Mayer's Hematoxylin followed by blueing in alkaline tap water or a 0.5% aqueous methyl green solution is recommended.
      7. Mount tissue sections in a water-based mounting medium. Air-dry blots and store protected from light.
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      152226 0

      Documentation

      AEC Substrate Buffer SDS

      Title

      Safety Data Sheet (SDS) 

      AEC Substrate Buffer Certificates of Analysis

      TitleLot Number
      152226
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision17-July-2007 RFH
      ApplicationImmunoblotting (see comments)
      Immunohistochemistry (see comments)
      DescriptionA diluent for use with 3-Amino-9-ethylcarbazole (AEC) (Cat. No. 152224), a precipitable peroxidase substrate for immunoblotting and immunohistochemical staining techniques, supplied as a 10X concentrate.
      FormLiquid
      FormulationIn 500 mM acetate buffer, 88 mM H₂O₂, proprietary stabilizers.
      PreservativeNone
      CommentsStable at 4°C, 18-26°C, and 37°C. When diluted the final pH is ~5.04. Functionality tested by performing immunohistologic stain using the final product.

      Suggested procedure for use of AEC in immunohistochemistry and immunoblotting:

      1. Complete all required incubations with antibodies and HRP-labeled probes, and all necessary wash steps.
      2. Prepare 1X AEC Substrate Buffer: Add 9 parts distilled water to 1 part 10X AEC Substrate Buffer, Cat. No. 152226 (0.5 M acetate buffer containing 88 mM H2O2). Mix well. The final pH of this solution will be ~5.0.
      3. Prepare 1X AEC Substrate Solution: Add 1 part 50X AEC Concentrate (Cat. No. 152224) to 50 parts 1X AEC Substrate Buffer (prepared in Step 2). Mix well.
      4. Following the final wash, completely cover tissue sections with 1X AEC Substrate Solution (prepared in Step 3) and incubate 10-30 min at room temperature. The same conditions apply for immunoblotting. Note: Variables associated with assay conditions will dictate the proper reaction times.
      5. After reaction is complete, wash tissue sections or blotting membranes thoroughly in distilled water.
      6. If tissue sections require a counterstain, Mayer's Hematoxylin followed by blueing in alkaline tap water or a 0.5% aqueous methyl green solution is recommended.
      7. Mount tissue sections in a water-based mounting medium. Air-dry blots and store protected from light.
      Storage +2°C to +8°C
      Do Not Freeze Ok to freeze
      Toxicity Standard Handling