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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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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
Catalogue 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