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Description
Overview
Precipitable peroxidase substrate for immunoblotting and immunohistochemical staining techniques. Produces a red end product that is soluble in alcohol. When used in immunohistochemistry, aqueous mounting media, and counterstains must be used. Directions included for use in each application.
Catalogue Number
152224
Brand Family
Calbiochem®
Synonyms
3-Amino-9-ethylcarbazole, 3AC
References
References
Okano, K., et al. 1992. Rinsho Byori 40, 93. Krenacs, T., et al. 1990. Histochem. J. 22, 530.
Immunoblotting (see comments) Immunohistochemistry (see comments)
Application Comments
Stable at 4°C and 18-26°C. Functionality determined by performing an 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 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
RTECS
FE3590000
Safety Information
R Phrase
R: 25
Toxic if swallowed.
S Phrase
S: 45
In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).
Product Usage Statements
Storage and Shipping Information
Ship Code
Ambient Temperature Only
Toxicity
Toxic
Hazardous Materials Attention:
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
Okano, K., et al. 1992. Rinsho Byori 40, 93. Krenacs, T., et al. 1990. Histochem. J. 22, 530.
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.
Revision
20-November-2008 JSW
Synonyms
3-Amino-9-ethylcarbazole, 3AC
Application
Immunoblotting (see comments) Immunohistochemistry (see comments)
Description
Precipitable peroxidase substrate for immunoblotting and immunohistochemical staining techniques supplied as a 50X concentrate (0.095 mol/l). To be used with AEC Substrate Buffer, Cat. No. 152226
Background
Precipitable peroxidase substrate for immunoblotting and immunohistochemical staining techniques. In the presence of horseradish peroxidase (HRP) and hydrogen peroxide, AEC is oxidized to a water-insoluble red product. AEC may be used as a substitute for diaminobenzidine (DAB) in HRP-based immunoassays. Although not as sensitive as DAB, the red deposit formed at the sites of AEC oxidation make this substrate useful where double labeling is desired. Since oxidized AEC is soluble in alcohols and xylene, aqueous-based mounting medium and counterstains must be used for immunohistochemical applications.
Form
Yellowish-brown to reddish solution
CAS number
132-32-1
RTECS
FE3590000
Chemical formula
C₁₄H₁₄N₂
Preservative
None
Comments
Stable at 4°C and 18-26°C. Functionality determined by performing an 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 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
Protect from light
+2°C to +8°C
Do Not Freeze
Ok to freeze
Special Instructions
Discard if a red or brown precipitate is evident.
Toxicity
Toxic
References
Okano, K., et al. 1992. Rinsho Byori 40, 93. Krenacs, T., et al. 1990. Histochem. J. 22, 530.
