281751 | DAB, Tetrahydrochloride, 50X Concentrate

281751
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      Overview

      Replacement Information

      Key Specifications Table

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      281751-10ML
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          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
          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.
          Biological Information
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          RTECSDV8753000
          Safety Information
          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.
          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).
          Packaging Information
          Transport Information
          Supplemental Information
          Specifications

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          281751

          References

          Reference overview
          Krueger, 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.

          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.

          Revision21-August-2007 JSW
          Synonyms3,3ʹ-Diaminobenzidine
          ApplicationImmunoblotting (see comments)
          Immunohistochemistry (see comments)
          DescriptionPeroxidase substrate that forms an insoluble, brown precipiate. Supplied as a 50X concentrate. Designed for use with DAB Substrate Buffer (Cat. No. 281753).
          BackgroundSince first introduced by Graham and Karnovsky numerous procedures for the use of DAB for detection of horseradish peroxidase (HRP)-labeled probes in histochemistry, immunohistochemistry, western blots and dot blots have been described. In the presence of horseradish peroxidase and hydrogen peroxide, DAB is oxidized to a brown polymer that is insoluble in most organic solvents. Thus, xylene-based mounting media may be used for immunohistochemical applications. Sites of HRP activity on tissue sections and blots appear as brown-orange deposits. The color can be modified and intensified by treatment with salts of silver, copper, nickel, cobalt and osmium. This DAB substrate preparation is a stable, convenient, liquid concentrate in a proprietary solvent. The stabilization system prevents formation of partially oxidized DAB, thus eliminating the nonspecific binding to other heme-containing proteins so often observed with powdered DAB preparations. The concentrate can be diluted in appropriate peroxide-containing buffers, providing the researcher with the capability of formulating any of the numerous published DAB reaction systems.
          FormBrown to dark brown solution
          FormulationSupplied as a 50X concentrate.
          CAS number7411-49-6
          RTECSDV8753000
          PreservativeNone
          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.
          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).
          Toxicity Toxic
          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.