DAB, 50X (CAS 7411-49-6)

5.0(3)

Alternate Names: sc-24982 also known as 3,3′ Diaminobenzidine Tetrahydrochloride; 3,3′-Diaminobenzidine-4HCl; DAB.4HCl

Application: DAB, 50X is 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: C₁₂H₁₄N₄•₄HCl

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

* Refer to Certificate of Analysis for lot specific data (including water content).

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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 :

n²⁰_D 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

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DAB, 50X (CAS 7411-49-6) Product Citations

See how others have used DAB, 50X (CAS 7411-49-6). Click on the entry to view the PubMed entry .

Citations 1 to 10 of 82 total

PMID: # 34394184 Chen, Z.|Chen, Z.|Xu, S.|Zhang, Q.| et al. 2021. Front Genet. 12: 674856.

PMID: # 30481381 Dasgupta, H. et al. 2018. J Surg Oncol.

PMID: # 30013653 Yan, T. et al. 2018. Oncol Lett. 16: 2591-2598.

PMID: # 29805628 Liu, ZG. et al. 2018. Oncol Lett. 15: 8935-8940.

PMID: # 30178275 Dasgupta, H. et al. 2018. Mol. Cell. Biochem.

PMID: # 27646721 Dasgupta, H. et al. 2017. Future Oncol. 13: 159-174.

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

Citations 1 to 10 of 82 total

In technical information I see Tris hydrochloride solution, pH 7.6 (SC-301951) - 1M as diluent but below in question section Tech supporter mentioned to dilute "with 100mM Tris-HCl pH 7.6". Where is true?

Asked by: jacki27

Thank you for your question. Since this is provided as 50X, you would dilute to 1x with 100mM Tris-HCl pH 7.6. If you have any further questions, please contact technical service.

Answered by: Technical Service 4

Date published: 2020-11-17

What is the concentration (mg/ml) of this product?

Asked by: lympho

Thank you for your question. The concentration for this product is 0.0694Mol/L. Here is a link to a sample CoA. http://datasheets.scbt.com/coa/coa.php?sc=sc-24982&lot=G1718

Answered by: Tech Service

Date published: 2019-09-26

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: 2020-02-09

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 <[email protected]>, 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 [email protected] if you continue to see the purple color with precipitate.

Answered by: Tech Service

Date published: 2020-12-24

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

Rated 5 out of 5 by Frontiers 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 qiushanmumeizi 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 MS 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

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