DAB, 50X A product used in HRP-based assays suitable for immunoblotting and immunohistochemical staining of nucleic acids and proteins

DAB, 50X (CAS 7411-49-6)

DAB, 50X | CAS 7411-49-6 is rated 5.0 out of 5 by 3.
  • y_2019, m_3, d_18, h_17
  • bvseo_bulk, prod_bvrr, vn_bulk_2.0.11
  • cp_1, bvpage1
  • co_hasreviews, tv_0, tr_3
  • loc_en_US, sid_24982, prod, sort_[SortEntry(order=SUBMISSION_TIME, direction=DESCENDING)]
  • clientName_scbt
  • bvseo_sdk, java_sdk, bvseo-3.2.0
  • CLOUD, getAggregateRating, 130ms
  • REVIEWS, PRODUCT
Synonym: 3,3' Diaminobenzidine Tetrahydrochloride
Application: A product used in HRP-based assays suitable for immunoblotting and immunohistochemical staining of nucleic acids and proteins
CAS Number: 7411-49-6
Molecular Weight: 360.11
Molecular Formula: C12H14N44HCl
* Refer to Certificate of Analysis for lot specific data (including water content).
Submit a review for this product and receive 15 CruzCredits

DAB (3-3′ diaminobenzidine tetrachloride) is suitable for immunoblotting and immunohistochemical staining of nucleic acids and proteins. In HRP-based assays, the buffer oxidizes to a brown polymer in the presence of HRP and hydrogen peroxide that is insoluble in most organic solvents; sites of HRP activity on tissue sections and blots appear as brown-orange deposits, which can be intensified by treatment with salts of silver, copper, nickel, cobalt and osmium.


References

1. Krueger, S.K., et al. 1999. Biotech. Histochem. 74: 105-109. PMID: 10333408
2. Gallyas, F., et al. 1982. J. Histochem. Cytochem. 30: 183-184. PMID: 7061820
3. Larrson, L.I. 1988. Immunocytochemistry: Theory and Practice. Boca Raton, FL. CRC Press, Inc.

Usage :
For Immunohistochemical:

1. To 50 parts of Tris hydrochloride solution, pH 7.6 (SC-301951), add 1 part of DAB, 50X and 1 part of 0.5% hydrogen peroxide aqueous solution. Mix well and protect from light.

1a. Hydrogen peroxide, 0.5%, prepared from 30% hydrogen peroxide (sc-203336) as follows: Dilute 0.167mL of 30% hydrogen peroxide to 10 mL with reagent grade water.

2. Completely cover tissue sections with this solution and incubate 5 - 15 minutes at room temperature.

3. After reaction is complete, wash sections thoroughly in reagent grade water.

4. Counterstain with hemotoxylin (sc-24973) if desired.

5. Dehydrate in graded alcohols to xylene or xylene substitutes.

6. Mount using xylene based mounting media (e.g., sc-45087).

For Blotting:

1. To 50 parts of Tris/maleic acid buffer, add 1 part of DAB Concentrate and 1 part of 0.5% hydrogen peroxide aqueous solution. Mix well and protect from light.

1a. Hydrogen peroxide, 0.5%, prepared from 30% hydrogen peroxide (sc-203336) as follows: Dilute 0.167mL of 30% hydrogen peroxide to 10 mL with reagent grade water.

1b. Tris/Maleic acid buffer prepared as follows: Dissolve 12.1g of tris base (sc-3715) in approximately 800mL of reagent grade water. Adjust pH to 6.0 with a 2 M solution of maleic acid (prepared with sc-250287). QS to 1 L with reagent grade water.


2. Completely cover membranes with this solution and incubate 5 - 30 minutes at room temperature.

3. After reaction is complete, wash membranes thoroughly in reagent quality water.

4. Air dry and store protected from light.

* Discard if there is precipitate in the solution prior to mixing with hydrogen peroxide. Discard if reagent is still purple after making solution with hydrogen peroxide.
Formulation :
50X concentrate
Appearance :
Solution
Physical State :
Liquid
Solubility :
Soluble in water (Dilutable). Insoluble in most organic solvents.
Storage :
Store at 4° C
Refractive Index :
n20D 1.77 (Predicted)
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
PubChem CID :
23892
MDL Number :
MFCD00012969
EC Number :
231-018-9
SMILES :
C1=CC(=C(C=C1C2=CC(=C(C=C2)N)N)N)N.Cl.Cl.Cl.Cl

Download SDS (MSDS)

Certificate of Analysis

Adobe Acrobat Reader is required to reliably view,
print and comment on PDF documents

DAB, 50X  Product Citations

See how others have used DAB, 50X. Click on the entry to view the PubMed entry .

Citations 1 to 10 of 59 total

PMID: # 28495142  Dong, Y. et al. 2017. J. Neuroimmunol. 307: 7-13.

PMID: # 26482544  Xing, R. et al. 2016. Scand. J. Immunol. 83: 64-71.

PMID: # 27008403  Fan, M. et al. 2016. Cell. Physiol. Biochem. 38: 1319-32.

PMID: # 27208794  Mukherjee, N. et al. 2016. Biochim. Biophys. Acta. 1862: 1472-84.

PMID: # 25465073  Maiti, GP. et al. 2015. Oral Surg Oral Med Oral Pathol Oral Radiol. 119: 202-12.

PMID: # 26500110  Zhou, R. et al. 2015. Experimental cell research. 339: 360-6.

PMID: # 26339260  Jafari, S. et al. 2014. Res Pharm Sci. 9: 453-61.

PMID: # 25543042  Wu, J. et al. 2014. Asian Pac. J. Allergy Immunol. 32: 316-20.

PMID: # 24656731  Zhang, L. et al. 2014. Biomaterials. 35: 4878-87.

PMID: # 23361078  Huang, QY. et al. 2013. Am J Forensic Med Pathol. 34: 68-71.

Citations 1 to 10 of 59 total

HOW TO PREPARE THE WORKING DAB SOLUTION FOR IHC BY USING DAB 50X SOLUTION??

Asked by: Pawan
Thank you for your question. This is provided as 50X. Dilute to 1x with 100mM Tris-HCl pH 7.6.
Answered by: Technical Support
Date published: 2018-11-30

It is necessary to add hydrogen peroxide to part ?. If so, what volume of hydrogen peroxide is necessary to add ?. Thank you.

Asked by: Pinatar84
Thank you for your message. Please contact our Technical Service by phone or email or by Live Chat.
Answered by: SCBT Heidelberg Support
Date published: 2018-11-27

I was wondering why do we need to discard the DAB solution if precipitates form. I didn't notice the warning before use. I coupled it with a secondary anti-body HRP. However, no band formation was seen for my western blot. Could this be the reason?

Asked by: David Dang
Thank you for your question. We recommend to discard the DAB solution if precipitates are observed prior to use. This is because the DAB should be in solution prior to mixing with the oxidizing agent in order for it to be effective. This is certainly one possibility for why you did not observe any bands after combining with your HRP conjugated secondary antibody, although there are also other possible contributing factors. Please contact our Technical Service Department and we are happy to assist you with troubleshooting your application. Please call us toll-free at (800) 457-3801 option 2, e-mail <scbt@scbt.com>, or use the live chat on our website.
Answered by: Technical Support
Date published: 2017-08-16

Hello, I bought thus product and when i tested it didn't work. I diluate dab as recommend in this page but solution continued purple , i teste the pH of tris and it was correct. What did i do wrong?

Asked by: Luã Macedo
Thank you for your question! The solution is fine purple, as long as there is not precipitate. What did you dilute the DAB in? The color should go away/lessen when you add Hydrogen Peroxide. Please contact your local distributor or email us at scbt@scbt.com if you continue to see the purple color with precipitate.
Answered by: Tech Service
Date published: 2017-05-29

What is the composition of this product?

Asked by: hawkeye11
This chemical, DAB, 50X (CAS 7411-49-6), is comprised of DAB, water, and an additive formulation that is non-toxic.
Answered by: Chemical Support 1
Date published: 2017-03-31
  • y_2019, m_3, d_16, h_16CST
  • bvseo_bulk, prod_bvqa, vn_bulk_2.0.11
  • cp_1, bvpage1
  • co_hasquestionsanswers, tq_5
  • loc_en_US, sid_24982, prod, sort_[SortEntry(order=LAST_APPROVED_ANSWER_SUBMISSION_TIME, direction=DESCENDING)]
  • clientName_scbt
  • bvseo_sdk, java_sdk, bvseo-3.2.0
  • CLOUD, getContent, 85ms
  • QUESTIONS, PRODUCT
Rated 5 out of 5 by from great This DAB is ready to use, VERY affordable price. The quality of the reagent is nothing but wonderful.
Date published: 2019-01-28
Rated 5 out of 5 by from Tech The product working very good. The technic support is professional and fast shipping
Date published: 2018-05-15
Rated 5 out of 5 by from Various publications cite DAB for its use in HRP Various publications cite DAB for its use in HRP-based assays suitable for immunoblotting and immunohistochemical staining of nucleic acids and proteins. -SCBT Publication Review
Date published: 2015-01-20
  • y_2019, m_3, d_18, h_17
  • bvseo_bulk, prod_bvrr, vn_bulk_2.0.11
  • cp_1, bvpage1
  • co_hasreviews, tv_0, tr_3
  • loc_en_US, sid_24982, prod, sort_[SortEntry(order=SUBMISSION_TIME, direction=DESCENDING)]
  • clientName_scbt
  • bvseo_sdk, java_sdk, bvseo-3.2.0
  • CLOUD, getReviews, 6ms
  • REVIEWS, PRODUCT

Santa Cruz Biotechnology, Inc. is a world leader in the development of products for the biomedical research market. Call us Toll Free at 1-800-457-3801.
Copyright © 2007-2019 Santa Cruz Biotechnology, Inc. All Rights Reserved. "Santa Cruz Biotechnology", and the Santa Cruz Biotechnology, Inc. logo, "Santa Cruz Animal Health", "San Juan Ranch", "Supplement of Champions", the San Juan Ranch logo, "Ultracruz", "Chemcruz", "Immunocruz", "Exactacruz", and "EZ Touch" are registered trademarks of Santa Cruz Biotechnology, Inc.
All trademarks are the property of their respective owners.