152226 | AEC Substrate Buffer

152226
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      Overview

      Replacement Information

      Key Specifications Table

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      152226-50ML
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      Available
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          Glass bottle 50 ml
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          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
          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

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          152226

          Brochure

          Title
          Antibody Sourcebook!" Second Edition
          Enzyme-Linked Detection Systems Technical Bulletin
          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