FAP expression in human tumor tissue was assessed using a pancancer TMA obtained from the University of Virginia. This TMA included 141 patients with 14 different types of cancer (bile duct, bladder, breast, colon, esophagus, stomach, liver, lung, ovary, oropharynx, pancreas, prostate, kidney, and uterus; 6–14 tumors per tissue type). Normal tissues present on the TMA were also evaluated (5–8 samples per tissue type). After deparaffinization and rehydration, heat-induced antigen retrieval (sodium citrate, 0.05% polysorbate 20, pH 6.0) was performed for 20 min using a vegetable steamer followed by quenching of endogenous peroxidase activity (3% hydrogen peroxide, 10 min). Primary antibody incubation with a 1:50 dilution of rabbit monoclonal anti-FAP α-[EPR20021] (ab207178; Abcam) was performed overnight at 4°C. Detection was performed using the ultraView Universal DAB Detection Kit (K3467; DAKO) per the manufacturer’s instructions. An experienced surgical pathologist (DWD) confirmed the histologic diagnoses and performed a immunohistochemistry analysis using a semiquantitative visual scoring system (0, negative staining; 1, weak staining; 2, strong staining).
Ultraview universal dab detection kit
Manufactured by Agilent Technologies
The UltraView Universal DAB Detection Kit is a lab equipment product designed for immunohistochemical detection of target proteins in tissue samples. It provides a chromogenic detection system using the 3,3'-Diaminobenzidine (DAB) substrate.
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Lab products found in correlation
2 protocols using ultraview universal dab detection kit
1
Immunohistochemical Analysis of FAP Expression
2
Immunohistochemical Analysis of Hepatic Cytolysis
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Three patients with persistent hepatic cytolysis underwent liver and digestive system biopsies.
In addition, one patient underwent skin biopsy twice for rashes. The biopsy samples were fixed in formalin and embedded in paraffin. Four-micrometer sections were generated and stained with hematoxylin-eosin-saffron. An immunohistochemical study was carried out using an automated immunohistochemistry system with antibodies against the following: CD20 (L26, Dako, 1/20e), CD3 (polyclonal rabbit, Dako, 1/100e), CD4 (4B12, Leica, 1/40e), CD8 (4B11, Leica, 1/50e), NLRP3 (HPA012878, Atlas antibodies, 1/400e), IL-1β (3A6, Cell Signaling, 1/100e), and NF-κB p65 (D14E12, Cell Signaling, 1/1500e). Briefly, 4-µm sections were prepared from paraffin blocks and placed on Superfrost plus slides. After antigen unmasking with target retrieval solution, citrate pH 6 or 9 was added according to the antibodies to be used, and the immunohistochemical study was carried out by an immunohistochemistry automaton (Ventana Benchmark XT). After incubation with the primary antibody (20 to 60 min), detection was carried out with an Ultraview Universal DAB Detection Kit (Dako) indirect detection kit according to the supplier's instructions.
A biopsy sample of chronic active colitis was used as an immunohistochemical control for some antibodies.
In addition, one patient underwent skin biopsy twice for rashes. The biopsy samples were fixed in formalin and embedded in paraffin. Four-micrometer sections were generated and stained with hematoxylin-eosin-saffron. An immunohistochemical study was carried out using an automated immunohistochemistry system with antibodies against the following: CD20 (L26, Dako, 1/20e), CD3 (polyclonal rabbit, Dako, 1/100e), CD4 (4B12, Leica, 1/40e), CD8 (4B11, Leica, 1/50e), NLRP3 (HPA012878, Atlas antibodies, 1/400e), IL-1β (3A6, Cell Signaling, 1/100e), and NF-κB p65 (D14E12, Cell Signaling, 1/1500e). Briefly, 4-µm sections were prepared from paraffin blocks and placed on Superfrost plus slides. After antigen unmasking with target retrieval solution, citrate pH 6 or 9 was added according to the antibodies to be used, and the immunohistochemical study was carried out by an immunohistochemistry automaton (Ventana Benchmark XT). After incubation with the primary antibody (20 to 60 min), detection was carried out with an Ultraview Universal DAB Detection Kit (Dako) indirect detection kit according to the supplier's instructions.
A biopsy sample of chronic active colitis was used as an immunohistochemical control for some antibodies.
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