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Diaminobenzidine substrate kit

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The Diaminobenzidine substrate kit is a laboratory reagent used for the detection and visualization of enzymatic activity in various biological samples. The kit contains the necessary components to perform a colorimetric reaction, resulting in the production of a brown, insoluble precipitate at the site of enzyme activity. This substrate is commonly used in immunohistochemistry and other related techniques.

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Lab products found in correlation

Ab2378, by Abcam (4 mentions) Ahr 002, by Alomone (4 mentions) Alexa uor 647, by Abcam (3 mentions) 6 diamidino 2 phenylindole dapi, by Thermo Fisher Scientific (2 mentions) Ab2057181, by Abcam (1 mentions)

5 protocols using diaminobenzidine substrate kit

1

Immunohistochemical Analysis of Intestinal Tissues

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Formalin-fixed, paraffin-embedded intestinal (cecal, colon and ileum) tissue section (4 μm) were incubated overnight at 4 °C with a primary antibody (Table 3) after antigen retrieval according to the manufacturer’s instructions.

Information of antibodies

Name of antibodyCompany name and Catalog no.Dilution factor
Anti-Mast Cell Tryptase antibody [AA1]abcam, ab23781:1000
Anti-IL-6 antibody [MP5-20F3]abcam, ab1911941:100
MUC2 AntibodyNovus, 312311:100
Goat Anti-Rabbit IgG H&L (HRP)abcam, ab2057181:1000
Goat Anti-Mouse IgG H&L (HRP)abcam ab970231:2500
Recombinant Human GDF11 protein (ab50159)abcam ab501591:1000
Samples were washed and subsequently incubated with secondary antibody for 45 min at RT (Table 3) Sections were counterstained either with the diaminobenzidine substrate kit (Nacalai, USA) followed by hematoxylin, or sections were stained with 6-diamidino-2-phenylindole (DAPI, Thermo Fisher, USA) for visualization of cell nuclei as previously described.
Conesa M.P., Blixt F.W., Peesh P., Khan R., Korf J., Lee J., Jagadeesan G., Andersohn A., Das T.K., Tan C., Di Gesu C., Colpo G.D., Moruno-Manchón J.F., McCullough L.D., Bryan R, & Ganesh B.P. (2023). Stabilizing histamine release in gut mast cells mitigates peripheral and central inflammation after stroke. Journal of Neuroinflammation, 20, 230.
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2

Immunohistochemical Characterization of Intestinal Mast Cells

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Formalin fixed, paraffin-embedded intestinal (cecal, ileum) tissue sections (4 μm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens (1) histamine receptor 2 (AHR-002, Alomone labs, Israel), (2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer’s instructions. Samples were washed and subsequently incubated with secondary antibody for 45 min at RT (Histofine simple stain anti-rabbit, 414341F, Nacalai, USA and Alexa fluor® 647, ab150131, Abcam, USA). Sections were counter-stained either with the diaminobenzidine substrate kit (Nacalai, USA) followed by hematoxylin, or sections were stained with 6-diamidino-2-phenylindole (DAPI, Thermo Fisher, USA) as previously described for visualization of cell nuclei.
Blasco M.P., Chauhan A., Honarpisheh P., Ahnstedt H., d’Aigle J., Ganesan A., Ayyaswamy S., Blixt F., Venable S., Major A., Durgan D., Haag A., Kofler J., Bryan R., McCullough L.D, & Ganesh B.P. (2020). Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation. Journal of Neuroinflammation, 17, 160.
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3

Immunohistochemical Analysis of Intestinal Tissue

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Formalin xed, para n-embedded intestinal (cecal, ileum) tissue sections (4 µm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens 1) histamine receptor 2 (AHR-002, Alomone labs, Israel), 2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer's instructions. Samples were washed and subsequently incubated with secondary antibody for 45 min at RT (Histo ne simple stain anti-rabbit, 414341F, Nacalai, USA and Alexa uor® 647, ab150131, Abcam, USA). Sections were counter-stained either with the diaminobenzidine substrate kit (Nacalai, USA) followed by haematoxylin or sections were stained with 6-diamidino-2-phenylindole (DAPI, Thermo scher, USA) as previously described for visualization of cell nuclei.
Blasco M.P., Chauhan A., Honarpisheh P., Ahnstedt H., d’Aigle J., Ganesan A., Ayyaswamy S., Blixt F.W., Venable S., Major A., Durgan D.J., Haag A.M., Kofler J., Bryan R.M., McCullough L.D., & Ganesh B.P. (2020). Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation.
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4

Immunohistochemical Analysis of Intestinal Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Formalin xed, para n-embedded intestinal (cecal, ileum) tissue sections (4 µm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens 1) histamine receptor 2 (AHR-002, Alomone labs, Israel), 2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer's instructions. Samples were washed and subsequently incubated with secondary antibody for 45 min at RT (Histo ne simple stain anti-rabbit, 414341F, Nacalai, USA and Alexa uor® 647, ab150131, Abcam, USA). Sections were counter-stained either with the diaminobenzidine substrate kit (Nacalai, USA) followed by haematoxylin or sections were stained with 6-diamidino-2-phenylindole (DAPI, Thermo scher, USA) as previously described for visualization of cell nuclei.
Blasco M.P., Chauhan A., Honarpisheh P., Ahnstedt H., d’Aigle J., Ganesan A., Ayyaswamy S., Blixt F.W., Venable S., Major A., Durgan D.J., Haag A.M., Kofler J., Bryan R.M., McCullough L.D., & Ganesh B.P. (2020). Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation.
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5

Immunohistochemical Analysis of Intestinal Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Formalin xed, para n-embedded intestinal (cecal, ileum) tissue sections (4 µm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens 1) histamine receptor 2 (AHR-002, Alomone labs, Israel), 2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer's instructions. Samples were washed and subsequently incubated with secondary antibody for 45 min at RT (Histo ne simple stain anti-rabbit, 414341F, Nacalai, USA and Alexa uor® 647, ab150131, Abcam, USA). Sections were counter-stained either with the diaminobenzidine substrate kit (Nacalai, USA) followed by haematoxylin or sections were stained with 6-diamidino-2-phenylindole (DAPI, Thermo scher, USA) as previously described for visualization of cell nuclei.
Blasco M.P., Chauhan A., Honarpisheh P., Ahnstedt H., d’Aigle J., Ganesan A., Ayyaswamy S., Blixt F.W., Venable S., Major A., Durgan D.J., Haag A.M., Kofler J., Bryan R.J., McCullough L.D., & Ganesh B.P. (2020). Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation.
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