Immunohistochemical Ki‐67 staining of 5 representative FTs was performed in sections from paraffin‐embedded tissue blocks with the avidin‐biotin‐peroxidase technique. Five micrometer‐thick sections were cut from the tissue specimens and placed on poly‐l ‐lysine‐coated glass slides. Sections were deparaffinized with xylene and rehydrated in graded alcohol. Endogenous peroxidase was blocked by immersing the sections in 0.1% hydrogen peroxidase in absolute methanol for 20 min. For antigen retrieval, tissue sections were heated in a pressure cooker in 10 mM citric acid monohydrate, pH 6.0 for 5 min, and then incubated with the primary antibody at room temperature. The primary antibody used was CONFIRM anti‐Ki‐67 (30–9) (Ventana Medical Systems, Tucson, AZ). Immunohistochemistry was performed with the Ventana BenchMark XT slide processing system and the iView detection kit (Ventana Medical Systems, Tucson, AZ). All slides were counterstained with hematoxylin, dehydrated, and mounted. Negative controls were performed by omitting the primary antibody.
Confirm anti ki 67 30 9
Manufactured by Roche
Sourced in United States
The CONFIRM anti-Ki-67 (30–9) is a laboratory diagnostic tool used to detect the presence and quantify the expression of the Ki-67 protein in tissue samples. Ki-67 is a cellular marker for proliferation and is widely used in the assessment of various types of cancer.
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Lab products found in correlation
I view detection kit, by Roche (1 mentions)
Benchmark xt platform, by Roche (1 mentions)
Ventana benchmark ultra, by Roche (1 mentions)
Optiview dab ihc detection kit, by Roche (1 mentions)
Cintec p16 histology, by Roche (1 mentions)
Optiview dab ihc detection kit, by Benchmark Scientific (1 mentions)
5 protocols using confirm anti ki 67 30 9
1
Immunohistochemical Ki-67 Analysis of FTs
2
Immunohistochemical Biomarker Evaluation
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Biomarker status were evaluated using pre-NAC and surgical FFPE tissue blocks by IHC. IHC staining of tumor tissues for ER (Confirm anti-ER (SP1), rabbit monoclonal antibody, Ventana Medical Systems), PR (Confirm anti-PR (1E2), rabbit monoclonal antibody, Ventana Medical Systems), human epidermal growth factor receptor 2 (HER2) (Ventana anti-HER2/neu (4B5), rabbit monoclonal antibody, Ventana Medical Systems), and Ki67 (Confirm anti-Ki67 (30–9), rabbit monoclonal antibody, Ventana Medical Systems) was performed using the automated Benchmark XT platform (Ventana Medical Systems) and according to the manufacturer’s recommendations. The cutoff value of ER- and PR-positive disease was no less than 1% positively stained nuclei in tumor tissues. HER2-positive status was defined as 3(+) by IHC or amplification confirmed by fluorescence in situ hybridization (FISH). Ki67 expression was divided into high expression group (≥ 15%) and low expression group (< 15%).
3
Quantifying Tumor Cell Proliferation
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Normal and neoplastic tissue proliferation was detected using a rabbit monoclonal primary antibody CONFIRM® anti-Ki-67 (30-9) (790-4286, Ventana Medical Systems, Tucson, Arizona, USA), which serves directly against the C-terminal portion of the Ki-67 antigen. The applied antibody for routine diagnostic IHC obtained FDA approval (510 k) for IVD (in vitro diagnostic) use. Brown nuclear (DAB) staining in tumor cells was identified as Ki-67 positive cells. The ARIOL imaging system (Applied Imaging Corp., San Jose, CA, USA) and light microscope quantified Ki-67 index.
4
Multiplex IHC Staining for Cervical Cancer
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IHC staining of p16, Ki‐67 and E4 for all biopsies was performed on in‐between sections using the Ventana BenchMark ULTRA automated slide stainer (Ventana Medical Systems, Roche, Tucson, AZ) at DDL Diagnostic Laboratory, Rijswijk, The Netherlands.19 , 20 IHC staining was performed after heat‐induced epitope retrieval with ULTRA Cell Conditioning Solution 1 (ULTRA CC1) and protease 3 (Roche, Basel, Switzerland) using mouse monoclonal antibodies against p16INK4a antigen (clone E6H4, CINtec, p16 Histology, Roche, Basel, Switzerland), rabbit monoclonal antibody against Ki‐67 [CONFIRM anti‐Ki‐67 (30‐9), Roche, Basel, Switzerland] and mouse monoclonal antibody against panHPV‐E4 antigen (SILgrade‐E4‐1 kit containing XR‐E4‐1 monoclonal antibody, Labo Biomedical Products B.V., Rijswijk, The Netherlands). The panHPV‐E4 antigen is at least reactive to HPV6, 11, 16, 18, 31, 33, 35, 39, 42, 43, 44, 45, 51, 52, 53, 56, 58, 59, 66, 67, 70 and 74; and skin HPV genotypes 27 and 57 (in part unpublished data).19 , 21 , 26 Reactivity was visualised using the OptiView DAB IHC Detection Kit for p16 and Ki‐67 detection, and the OptiView DAB IHC Detection Kit with OptiView Amplification Kit for HPV‐E4 detection (Roche, Tucson, AZ).
5
Immunohistochemical Analysis of Ki67
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The Bouin-fixed paraffin-embedded biopsies and PFA-fixed paraffin-embedded rat testes were subjected to immunohistochemical analysis using the VENTANA Roche automated immunostainer (Roche, Mannheim, Germany). The VENTANA primary Ki67 antibody (Roche) has been developed for use on BenchMark IHC/ISH instruments in conjunction with VENTANA detection kits and accessories. Tissue sections were deparaffinized and processed to unmask the antigenic sites. The CONFIRM anti-Ki-67 (30-9, Roche) Rabbit Monoclonal Primary Antibody was used. The sections were stained with OptiView DAB IHC Detection Kit in BenchMark instruments (Method GX, Roche).
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