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Dab reaction

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DAB (3,3'-Diaminobenzidine) is a chromogenic substrate used for immunohistochemical detection and visualization of target proteins in biological samples. It produces a brown-colored precipitate at the site of the enzyme-substrate reaction, allowing for the identification and localization of the target proteins within the sample.

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Lab products found in correlation

Hematoxylin, by Thermo Fisher Scientific (4 mentions) Abc substrate kit, by Vector Laboratories (4 mentions) Brdu antibody, by BD (2 mentions) γh2ax, by Cell Signaling Technology (2 mentions) Cd45r, by Abcam (2 mentions) Ab45977, by Abcam (1 mentions) Bx61 microscope, by Olympus (1 mentions) Ki 67 antibody ab15580, by Abcam (1 mentions) Cyclin d3 dcs22, by Cell Signaling Technology (1 mentions) Prb s780, by Cell Signaling Technology (1 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)

5 protocols using dab reaction

1

Immunohistochemical Analysis of Cerebral Artery

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Tissues were collected from CA and superficial temporal artery (STA) from 12 patients who underwent microsurgical clipping of CA. Samples were fixed in paraformaldehyde, and embedded in paraffin. Following antigen retrieval (by microwaving in pH6.0 citrate buffer), sections were incubated with anti-MPO antibody (ab45977, Abcam), followed by HRP-conjugated reagent (Boost, #8114, Cell Signaling) with DAB reaction (Vector Labs). Slides were stained with hematoxylin, dehydrated, and permanently mounted. Images were taken at 200x magnification with an Olympus BX61 microscope.
Chu Y., Wilson K., Gu H., Wegman-Points L., Dooley S.A., Pierce G.L., Cheng G., Pena Silva R.A., Heistad D.D, & Hasan D. (2015). Myeloperoxidase Is Increased in Human Cerebral Aneurysms and Increases Formation and Rupture of Cerebral Aneurysms in Mice. Stroke; a journal of cerebral circulation, 46(6), 1651-1656.
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2

Immunohistochemical Analysis of Lymphoma Xenografts

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Lymphoma tissues from xenograft were fixed with 4%PFA, dehydrated, and embedded in paraffin followed by sectioned with a microtome. H&E sections and tissue microarray slides (US Biomax, Inc.) were blocked with 10% goat serum, incubated with the primary antibodies and subsequently incubated with biotinylated antibodies. Signal was developed with ABC substrate kit (Vector) followed by DAB reaction (Vector), and counterstained with Hematoxylin (Thermo Scientific). The Ki-67 antibody (ab15580) was purchased from Abcam. Fbxl8 antibody (NBP2-34012) was obtained from Novus Biologicals. Cyclin D3 (DCS22) and pRb (S780) were purchased from Cell Signaling Technology and Santa Cruz, respectively. The IHC score in each core was defined by following formula Intensity = (staining positive population (1 to 3) × staining intensity (1 to3)). The IHC scores 1–2, 3–5, and 6–9 were defined as expression of low, medium, and high, respectively.
Yoshida A., Choi J., Jin H.R., Li Y., Bajpai S., Qie S, & Diehl J.A. (2020). Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3. Oncogene, 40(2), 292-306.
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3

Immunohistochemical Analysis of Cell Signaling

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Tissues were fixed in 4% paraformaldehyde, deparaffinized and rehydrated in gradient ethanol. Sections were blocked with 10% goat serum and incubated with the primary antibodies against the following: P-S6 (S235/236), P-Akt (S473), Ki67 and pRb (S780). For immunohistochemistry, sections were incubated with biotinylated rabbit antibodies and developed with ABC substrate kit (Vector) followed by DAB reaction (Vector), and counterstained with Hematoxylin (Thermo Scientific). For immunofluorescence, sections were incubated with Alex Fluor 488 or Alexa Fluor 546-conjugated goat anti-mouse/rabbit antibodies (Life technologies), and mounted with ProLong Gold antifade reagent with DAPI (Invitrogen).
Yoshida A., Lee E.K, & Diehl J.A. (2016). Induction of therapeutic senescence in vemurafenib-resistant melanoma by extended Inhibition of CDK4/6. Cancer research, 76(10), 2990-3002.
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4

Immunohistochemical Analysis of Tumor Samples

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Tumors from mice were fixed with 4% PFA, dehydrated, and embedded in paraffin followed by sectioned with a microtome. Sections were blocked with 10% goat serum, incubated with the primary antibodies, and subsequently incubated with biotinylated antibodies. Signal was developed with ABC substrate kit (Vector) followed by DAB reaction (Vector), and counterstained with Hematoxylin (Thermo Scientific). Antibodies for p-Rb (Ser780), p-Chk1 (Ser345), Pten, p-Akt (Ser473), Akt, and γH2AX were obtained from Cell Signaling Technology. Antibodies for Ki-67 (ab15580), p-Chk2 (Thr68), CD3 and CD45R were purchased from Abcam. BrdU antibody was obtained from BD Biosciences. Cyclin D1 (72–13G) was generated in our laboratory.
CORRECTION : Volume 117: 1401–1410, 1998. (1998). Plant Physiology, 118(3), 1103.
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5

Immunohistochemical Analysis of Tumor Samples

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Tumors from mice were fixed with 4% PFA, dehydrated, and embedded in paraffin followed by sectioned with a microtome. Sections were blocked with 10% goat serum, incubated with the primary antibodies, and subsequently incubated with biotinylated antibodies. Signal was developed with ABC substrate kit (Vector) followed by DAB reaction (Vector), and counterstained with Hematoxylin (Thermo Scientific). Antibodies for p-Rb (Ser780), p-Chk1 (Ser345), Pten, p-Akt (Ser473), Akt, and γH2AX were obtained from Cell Signaling Technology. Antibodies for Ki-67 (ab15580), p-Chk2 (Thr68), CD3 and CD45R were purchased from Abcam. BrdU antibody was obtained from BD Biosciences. Cyclin D1 (72–13G) was generated in our laboratory.
Yoshida A., Phillips-Mason P., Tarallo V., Avril S., Koivisto C., Leone G, & Diehl J.A. (2022). Non-phosphorylatable cyclin D1 mutant potentiates endometrial hyperplasia and drives carcinoma with Pten loss. Oncogene, 41(15), 2187-2195.
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