BAP1 immunocytochemistry was applied on 4 μm thick deparaffinized sections using Poly-L-Lysin slides from blocks containing sufficient tissue. IHC staining was performed in accordance with the manufacturer’s instructions. Automatic IHC staining device (Dako Omnis Automated IHC/ISH Staining system) was used in the study. BAP1 polyclonal antibody (BRCA1-associated Protein 1[C4], Catalog Number MC0136, Medaysis) was used at a 1:50 dilution.
Omnis automated ihc ish staining system
Manufactured by Agilent Technologies
The Omnis Automated IHC/ISH Staining system is a laboratory instrument designed for automated immunohistochemistry (IHC) and in situ hybridization (ISH) staining. It performs sample preparation, reagent dispensing, and incubation steps to facilitate the staining process for clinical and research applications.
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Lab products found in correlation
3 protocols using omnis automated ihc ish staining system
1
Immunohistochemical Detection of BAP1
2
Immunohistochemical Analysis of FoxP3+ Lymphocytes in HPV-Associated Cancers
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The paraffin block that best reflects the morphology was selected, and immunohistochemical studies were performed in accordance with the manufacturer’s instructions. Automatic immunohistochemistry staining device (Dako Omnis Automated IHC/ISH Staining System) was used in the study. FoxP3 (Abcam, clone 236A/E7) and p16/INK4a (Medaysis, clone G175-405) were administered. According to the p16 staining results, the cases were grouped as HPV-associated and independent. Cell counts were made in areas with the highest concentration of FoxP3 positive lymphocytes. Positive cells detected in tumor stroma and within tumoral cell groups were counted separately. Counting was done in 4 high magnification fields and the average cell number was determined. It was investigated whether clinicopathological features such as p16 staining status, presentation complaints of the cases, tumor size and localization, and treatment method were statistically different in tumors with low and high FoxP3 positive lymphocyte counts. It was studied whether the recurrence status and survival times changed statistically in these groups.
3
Immunohistochemical Analysis of NS Expression in Tumor Samples
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Paraffin blocks that best reflected tumor morphology were studied via immunohistochemical staining for NS (ab70346 -Abcam, Eugene, OR, USA). Sections with a thickness of 4 μm were obtained from these paraffin blocks, and slides were deparaffinized. Immunoperoxidase staining was then completed using an automatic staining machine (the Dako Omnis Automated IHC/ISH staining system) in accordance with the manufacturer's instructions. Chromogenic diaminobenzidine (DAB) was used for signal detection, and cells were counterstained with Harris hematoxylin. Negative controls were incubated with the same concentration of immunoglobulin (IgG1; Dako, Ely, UK) instead of the primary antibody. The positive controls were testicular seminoma specimens. Only nuclear staining was evaluated; cytoplasmic staining was not considered. The level of expression was assessed by the percentage and the intensity of the immunohistochemical staining. Pannuclear strong staining was classified as severe positive, while spot or mild nuclear staining was regarded as weak positive. The expression of NS compared with the Ki-67 proliferation index (routinely determined in cases by immunohistochemical method), tumor grade, and clinicopathological features.
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