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3 3 diaminobenzidine chromogen dab

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3,3'-Diaminobenzidine chromogen (DAB) is a widely used substrate for the visualization of enzymatic activity in immunohistochemistry and other molecular biology applications. It produces a brown precipitate at the site of the enzyme-catalyzed reaction, enabling the localization and detection of target analytes.

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Lab products found in correlation

Novolink polymer, by Leica (2 mentions) 24 pax 5, by BD (2 mentions) B220 antibody clone ra3 6b2, by BD (2 mentions) Novocastra post primary, by Leica (2 mentions)

Spelling variants of this product

3,3′-diaminobenzidine (13 citations) DAB chromogen (5 citations) 3,3′-Diaminobenzidine (DAB; (3 citations)

2 protocols using 3 3 diaminobenzidine chromogen dab

1

Immunohistochemical Profiling of ILC3 in Lung

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Immunohistochemical staining of human, NHP and mouse formalin-fixed paraffin-embedded (FFPE) lung sections were initially dewaxed in xylene prior to hydrating with decreasing graded alcohol and methanol passages. Antigen was retrieved via heat treatment in 92°C and EDTA buffer pH 8. Tissue staining with RORγt (Clone 6F3.1, Millipore for mouse; clone Q31–378, BD Bioscience for NHP and human), CD3 (clone SP7, Thermofisher for human, NHP and mouse) or PAX5 (Clone 24/Pax-5, BD Pharmingen, for human and NHP) or B220 antibody (clone RA3–6B2, BD Pharmingen) was performed for one hour in a humid chamber. Tissues were washed in Tris buffered saline pH7.4–7.6 prior to incubation with secondary antibody (Novocastra Post Primary, Leica) and polymer (Novolink Polymer, Leica). To develop the reaction, tissues were incubated with 3,3’-Diaminobenzidine chromogen (DAB, Leica). Singly stained sections (PAX5, B220) were incubated with DAB for 5 minutes and tissues receiving double staining (RORγt and CD3) were incubated overnight. Tissues were counterstained with haematoxylin and rinsed in water. All tissues were mounted with coverslips using glycerol mounting medium. CD3−RORγt+ ILC3 were quantified in the slides. Images were analyzed manually by counting the number of ILC3 cells per field. The analysis was done in a blinded manner.
Ardain A., Domingo-Gonzalez R., Das S., Kazer S.W., Howard N.C., Singh A., Ahmed M., Nhamoyebonde S., Rangel-Moreno J., Ogongo P., Lu L., Ramsuran D., de la Luz Garcia-Hernandez M., Ulland T., Darby M., Park E., Karim F., Melocchi L., Madansein R., Dullabh K.J., Dunlap M., Marin-Agudelo N., Ebihara T., Ndung’u T., Kaushal D., Pym A.S., Kolls J.K., Steyn A., Zúñiga J., Horsnell W., Yokoyama W., Shalek A.K., Kløverpris H.N., Colonna M., Leslie A, & Khader S.A. (2019). Group 3 innate lymphoid cells mediate early protective immunity against tuberculosis. Nature, 570(7762), 528-532.
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2

Immunohistochemical Profiling of ILC3 in Lung

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunohistochemical staining of human, NHP and mouse formalin-fixed paraffin-embedded (FFPE) lung sections were initially dewaxed in xylene prior to hydrating with decreasing graded alcohol and methanol passages. Antigen was retrieved via heat treatment in 92°C and EDTA buffer pH 8. Tissue staining with RORγt (Clone 6F3.1, Millipore for mouse; clone Q31–378, BD Bioscience for NHP and human), CD3 (clone SP7, Thermofisher for human, NHP and mouse) or PAX5 (Clone 24/Pax-5, BD Pharmingen, for human and NHP) or B220 antibody (clone RA3–6B2, BD Pharmingen) was performed for one hour in a humid chamber. Tissues were washed in Tris buffered saline pH7.4–7.6 prior to incubation with secondary antibody (Novocastra Post Primary, Leica) and polymer (Novolink Polymer, Leica). To develop the reaction, tissues were incubated with 3,3’-Diaminobenzidine chromogen (DAB, Leica). Singly stained sections (PAX5, B220) were incubated with DAB for 5 minutes and tissues receiving double staining (RORγt and CD3) were incubated overnight. Tissues were counterstained with haematoxylin and rinsed in water. All tissues were mounted with coverslips using glycerol mounting medium. CD3−RORγt+ ILC3 were quantified in the slides. Images were analyzed manually by counting the number of ILC3 cells per field. The analysis was done in a blinded manner.
Ardain A., Domingo-Gonzalez R., Das S., Kazer S.W., Howard N.C., Singh A., Ahmed M., Nhamoyebonde S., Rangel-Moreno J., Ogongo P., Lu L., Ramsuran D., de la Luz Garcia-Hernandez M., Ulland T., Darby M., Park E., Karim F., Melocchi L., Madansein R., Dullabh K.J., Dunlap M., Marin-Agudelo N., Ebihara T., Ndung’u T., Kaushal D., Pym A.S., Kolls J.K., Steyn A., Zúñiga J., Horsnell W., Yokoyama W., Shalek A.K., Kløverpris H.N., Colonna M., Leslie A, & Khader S.A. (2019). Group 3 innate lymphoid cells mediate early protective immunity against tuberculosis. Nature, 570(7762), 528-532.
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