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281751 DAB, Tetrahydrochloride, 50X Concentrate

This DAB, Tetrahydrochloride, 50X Concentrate is validated for use in Immunoblotting, Immunohistochemistry.

DAB, Tetrahydrochloride, 50X Concentrate MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: 3,3ʹ-Diaminobenzidine

281751
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Overview

Products

Catalogue NumberPackaging Qty/Pack
281751-10MLCN Glass bottle 10 ml
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Description
OverviewProduces a brown alcohol-insoluble end product. Also useful as a stain for myelin in glutaraldehyde-fixed sections. Supplied as a 50X concentrate.
Catalogue Number281751
Brand Family Calbiochem®
Synonyms3,3ʹ-Diaminobenzidine
References
ReferencesKrueger, S.K., et al. 1999. Biotech. Histochem. 74, 105.
Larrson, L.I. 1988. Immunocytochemistry: Theory and Practice, CRC Press, Inc., Boca Raton, FL.
Gallyas, F., et al. 1982. J. Histochem. Cytochem. 30, 183.
Hsu, S.M. and Soban, E. 1982. J. Histochem. Cytochem. 30, 1079.
Lewis, P.R. and Knight, D.P. 1977. Staining Methods for Sectioned Material, In Practical Methods in Electron Microscopy, A.M. Glauert, ed. North-Holland, Amsterdam.
Graham, R.C., Jr. and Karnovsky, M.J. 1966. J. Histochem. Cytochem. 14, 291.
Product Information
CAS number7411-49-6
FormBrown to dark brown solution
FormulationSupplied as a 50X concentrate.
PreservativeNone
Quality LevelMQ100
Applications
Application NotesImmunoblotting (see comments)
Immunohistochemistry (see comments)
Application CommentsStable at 4°C and 18-26°C.

Suggested Procedure for Immunohistochemical Staining Using DAB:

1. Complete all required incubations with antibodies and HRP-labeled probes, and all necessary wash steps.
2. Prepare 1X DAB Substrate Buffer: Add 9 parts distilled water to 1 part 10X DAB Substrate Buffer (Cat. No. 281753). Mix well. [Note: 1 M Tris-HCl, pH 7.6 may be substituted for Cat. No. 281753, such that the final (1X) concentration is 100 mM Tris-HCl, pH 7.6.
3. Prepare 1X DAB Substrate Solution: Add 1 part 50X DAB Concentrate and 1 part 0.5% H2O2 to 50 parts 1X DAB Substrate Buffer (prepared in Step 2). Mix well and protect from light.
4. Following the final wash, completely cover tissue sections with the 1X DAB Substrate Solution (prepared in Step 3) and incubate 5-15 min at room temperature. Note: Variables associated with assay conditions will dictate the proper reaction time.
5. After reaction is complete, wash tissue sections thoroughly in distilled water.
6. Counterstain with hematoxylin if desired.
7. Dehydrate in graded alcohols, xylene, or xylene substitutes.
8. Mount tissue sections using xylene-based mounting media.

Suggested Procedure for Immunoblot Staining with DAB:

1. Complete all required incubations with antibodies and HRP-labeled probes, and all necessary wash steps.
2. Prepare 1X DAB Substrate Buffer: Add 9 parts distilled water to 1 part 10X DAB Substrate Buffer (Cat. No. 281753). Mix well. [Note: 1 M Tris-HCl, pH 7.6 may be substituted for Cat. No. 281753, such that the final (1X) concentration is 100 mM Tris-HCl, pH 7.6.]
3. Prepare 1X DAB Substrate Solution: Add 1 part 50X DAB Concentrate and 1 part 0.5% H2O2 to 50 parts 1X DAB Substrate Buffer (prepared in Step 2). Mix well and protect from light.
4. Following the final wash, completely cover membranes with 1X DAB Substrate Solution (prepared in Step 3). Incubate 5-30 min at room temperature.
Note: Variables associated with assay conditions will dictate the proper reaction time.
5. After reaction is complete, wash membranes thoroughly in distilled water.
6. Air-dry membranes and store protected from light.
Safety Information according to GHS
RTECSDV8753000
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.
Storage +2°C to +8°C
Do not freeze Ok to freeze
Special InstructionsDiscard if a precipitate forms or if reagent is purple (the purple material is a DAB oxidation product that binds avidly to heme-containing proteins).
Global Trade Item Number
Catalogue Number GTIN
281751-10MLCN 04055977197808