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Rck108

Manufactured by Agilent Technologies
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Sourced in United States, Denmark

The RCK108 is a laboratory equipment product offered by Agilent Technologies. It serves as a rack system designed to accommodate various Agilent instruments and accessories. The core function of the RCK108 is to provide a modular and organized platform for housing and integrating Agilent's analytical instruments within a laboratory setting.

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Lab products found in correlation

Benchmark ultra automated slide stainer, by Roche (2 mentions) Halo image analysis platform, by Indica Labs (2 mentions) Nanozoomer xr, by Hamamatsu Photonics (2 mentions) Ov tl 12 30, by Agilent Technologies (2 mentions) Ventana benchmark ultra, by Roche (1 mentions) Cryo cut microtome, by Leica Biosystems (1 mentions) Leica bond 3 autostainer, by Leica Biosystems (1 mentions) Bond polymer refine detection kit, by Leica Biosystems (1 mentions) Ab92551, by Abcam (1 mentions) Ab24643, by Abcam (1 mentions) Ab7800, by Abcam (1 mentions) Envision dual link system hrp, by Agilent Technologies (1 mentions) Cellient automated cell block system, by Hologic (1 mentions) Benchmark ultra immunostainer, by Roche (1 mentions)

9 protocols using rck108

1

Detection of Lymph Node Micrometastases

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Formalin-fixed, paraffin-embedded tissue blocks of the lymph nodes were retrieved from the tissue archives of the departments of pathology in the AMC and UMCG. Four new levels of each lymph node with a distance of 250 µm between the levels were investigated. Two 4 µm thick sections of each level were cut serially. One section was stained with H&E and one with an antibody against K19 (dilation 1:100, clone RCK108; Dako, Glostrup, Denmark). All staining procedures were performed in the UMCG. The K19 staining was performed using a Ventana Benchmark Ultra automated stainer (Ventana Medical Systems, Tucson, AZ, USA) after pretreatment with protease.
Micrometastases were defined as cells detected by immunostaining with morphologic features of adenocarcinoma. H&E and K19 immunolabeled slides were investigated by two experienced pathologists (ASHG and JJD) blinded to the demographic and clinicopathologic features of patients.
Mantel H.T., Wiggers J.K., Verheij J., Doff J.J., Sieders E., van Gulik T.M., Gouw A.S, & Porte R.J. (2015). Lymph Node Micrometastases are Associated with Worse Survival in Patients with Otherwise Node-Negative Hilar Cholangiocarcinoma. Annals of Surgical Oncology, 22, 1107-1115.
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2

Quantification of CK19-Positive Cancer Cells in COD

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We performed immunohistochemical (IHC) labeling for cytokeratin 19 (CK19) on one representative slide per case with COD. Formalin-fixed, paraffin-embedded blocks were sectioned at 5 μm, deparaffinized, and subjected to antigen retrieval. Sections were then immunolabeled for CK19 (mouse monoclonal clone RCK108; 1:100 dilution; Dako, Denmark) using the BenchMark Ultra automated slide stainer (Ventana Medical Systems, AZ). We then scanned all the IHC slides at 20 × magnification using NanoZoomer XR (Hamamatsu Photonics, Japan). Regions of COD and carcinoma in stroma were separately annotated on each scanned IHC slide while concurrently reviewing the cellular morphology on the matched H&E slide using a microscope. This approach allowed distinction of normal ductal cells and PDAC, both of which are labeled by CK19 but have a strikingly different morphology on IHC and H&E stains. The number of CK19-positive cancer cells in each region was quantified using the HALO Image Analysis Platform (Indica labs, NW). Non-neoplastic CK19-positive epithelium was excluded from this quantification based on morphology.
Fujikura K., Hutchings D., Braxton A.M., Zhu Q., Laheru D.A., Hruban R.H., Thompson E.D, & Wood L.D. (2020). Intraductal Pancreatic Cancer is Less Responsive Than Cancer in the Stroma to Neoadjuvant Chemotherapy. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 33(10), 2026-2034.
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3

Rapid Immunohistochemistry for Frozen Tissue

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Fresh frozen tissue in OCT compound was sectioned with Cryo-cut Microtome (Leica Biosystems, Newcastle Upon Tyne, UK) in 3–4 μm thickness, placed on silane coated slide, and let dry. The slide was then stained for rapid immunohistochemistry in LEICA BOND-III Autostainer using Bond Polymer Refine Detection kit (Leica Biosystems). Briefly, the dry slide was fixed in 4% paraformaldehyde for 1 minute, immersed in peroxide block for 2 minutes to endogenous peroxidase blocking, washed and then applied with primary antibody for 4 minutes. After washing with Bond Wash solution, the slide was sequentially applied with post primary agent for 2 minutes and polymer for 2 minutes with washings in-between. The antibodies used were CK19 (1:80, RCK108, mouse monoclonal, DAKO, Carpinteria, CA, USA) and CD56 (1:50, 123C3, mouse monoclonal, DAKO). They were detected with 3,3’-diaminobenzidine (DAB) chromogen and DAB enhancer and counterstained with hematoxylin. The entire process takes roughly about 30 minutes.
Pyo J.Y., Choi S.E., Shin E., Koo J, & Hong S. (2017). The Intraoperative Immunohistochemical Staining of CD56 and CK19 Improves Surgical Decision for Thyroid Follicular Lesions. Journal of Pathology and Translational Medicine, 51(5), 463-470.
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4

Histological Scoring and Immunohistochemistry

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The paraffin-embedded tissues stained with H&E were scored in a blinded manner according to Ishak scoring system by a single pathologist. The degree of necroinflammatory activity and the stage of fibrosis were scored 0–18 or 0–6 respectively in the non-tumor specimen according to Ishak et al.[19 (link)]. Median values were used as a cut-off in subsequent analyses. Immunohistochemistry was carried out according to appropriate protocols [20 (link)]. The primary antibody used was mouse monoclonal anti-CK19 (1:100, clone RCK108, Dako), Blank controls were treated identically except that the primary antibodies were omitted.
Xu M., Xie F., Qian G., Jing Y., Zhang S., Gao L., Zheng T., Wu M., Yang J, & Wei L. (2014). Peritumoral ductular reaction: a poor postoperative prognostic factor for hepatocellular carcinoma. BMC Cancer, 14, 65.
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5

Comprehensive Cytokeratin Immunostaining Protocol

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Immunostaining was performed on cytokeratins (CK) 1, 10, 13, 14, 16, 18, and 19 using the Envision Dual Link System HRP (Dako, K4061). Cells were incubated with mouse anti-CK10 (1:100, DEK-10, BioGenex, Fremont, California, USA), anti-CK16 (1:300, ab8741, Abcam), anti-CK18 (1:1500, DC10, Dako), anti-CK19 (1:2000, RCK108, Dako), rabbit anti-CK1 (1:200, ab24643, Abcam), anti-CK13 (1:100, ab92551, Abcam), and anti-CK14 (1:2000, ab7800, Abcam) followed by the polymer-link method. The slides were scanned into high-resolution images using an Aperio ScanScope XT (Aperio Technology, Inc), Vista, CA). The images were then viewed in an Image Scope (Aperio Technology, Inc) and immunostaining was quantified using the Positive Pixel Count algorithm, v9 (Aperio).
Rodrigues M.F., de Oliveira Rodini C., de Aquino Xavier F.C., Paiva K.B., Severino P., Moyses R.A., López R.M., DeCicco R., Rocha L.A., Carvalho M.B., Tajara E.H, & Nunes F.D. (2014). PROX1 Gene is Differentially Expressed in Oral Cancer and Reduces Cellular Proliferation. Medicine, 93(28), e192.
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6

Cell Block Immunohistochemistry Protocol

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Cells were processed into formalin-fixed paraffin-embedded tissue blocks using the Cellient Automated Cell Block System (Hologic, Bedford, MA, USA) and processed for immunohistochemistry as described above. In addition, coated and non-coated cells were stained for K19 (1:50, Dako, RCK108) using Envision Flex anti-mouse secondary (Dako). Visualization was done using 3-amino-9-ethylcarbazole, followed by a hematoxylin counterstaining.
Govaere O., Petz M., Wouters J., Vandewynckel Y.P., Scott E.J., Topal B., Nevens F., Verslype C., Anstee Q.M., Van Vlierberghe H., Mikulits W, & Roskams T. (2017). The PDGFRα-laminin B1-keratin 19 cascade drives tumor progression at the invasive front of human hepatocellular carcinoma. Oncogene, 36(47), 6605-6616.
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7

Quantification of CK19-Positive Cancer Cells in COD

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We performed immunohistochemical (IHC) labeling for cytokeratin 19 (CK19) on one representative slide per case with COD. Formalin-fixed, paraffin-embedded blocks were sectioned at 5 μm, deparaffinized, and subjected to antigen retrieval. Sections were then immunolabeled for CK19 (mouse monoclonal clone RCK108; 1:100 dilution; Dako, Denmark) using the BenchMark Ultra automated slide stainer (Ventana Medical Systems, AZ). We then scanned all the IHC slides at 20 × magnification using NanoZoomer XR (Hamamatsu Photonics, Japan). Regions of COD and carcinoma in stroma were separately annotated on each scanned IHC slide while concurrently reviewing the cellular morphology on the matched H&E slide using a microscope. This approach allowed distinction of normal ductal cells and PDAC, both of which are labeled by CK19 but have a strikingly different morphology on IHC and H&E stains. The number of CK19-positive cancer cells in each region was quantified using the HALO Image Analysis Platform (Indica labs, NW). Non-neoplastic CK19-positive epithelium was excluded from this quantification based on morphology.
Fujikura K., Hutchings D., Braxton A.M., Zhu Q., Laheru D.A., Hruban R.H., Thompson E.D, & Wood L.D. (2020). Intraductal Pancreatic Cancer is Less Responsive Than Cancer in the Stroma to Neoadjuvant Chemotherapy. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 33(10), 2026-2034.
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8

Immunohistochemical analysis of liver samples

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Specimens of liver were fixed in 4% acetic formalin, embedded in paraffin, cut into 4‐micron sections, stained with hematoxylin and eosin (H&E), and immunostained with antibodies against cytokeratin (CK) 7 (monoclonal mouse anti‐human, OV‐TL12/30, ready to use; Agilent, Santa Clara, CA, USA) and CK19 (monoclonal mouse anti‐human, RCK108, ready to use; Agilent). Immunostaining with anti‐MYO5B antibodies (N‐term human MYO5B, Ab190096; Abcam, Cambridge, UK) and anti‐BSEP monoclonal antibody (mouse anti‐human, F‐6, sc‐74500; Santa Cruz Biotechnology, Dallas, TX, USA) were also performed. Specimens from patients and normal controls were stained together on the same slide when performing immunostaining.
For a diagnosis‐blinded review of BSEP (ABCB11) immunostaining, unstained slides were sent to B.S. Slides were subjected to heat‐induced antigen retrieval (CC1; Ventana Medical Systems, Tucson, AZ) and immunostained with rabbit polyclonal anti‐BSEP antibody (HPA 19035, 1:2,000 dilution; Sigma‐Aldrich, Taufkirchen, Germany), with diaminobenzidine as chromogen and hematoxylin as counterstain, using a Benchmark Ultra Immunostainer (Ventana). Normal human liver was used as a positive control. Slides were reviewed by two independent pathologists (A.S.K. and B.S.) with no knowledge of associated clinical or genetic information.
Qiu Y., Gong J., Feng J., Wang R., Han J., Liu T., Lu Y., Li L., Zhang M., Sheps J.A., Wang N., Yan Y., Li J., Chen L., Borchers C.H., Sipos B., Knisely A.S., Ling V., Xing Q, & Wang J. (2017). Defects in myosin VB are associated with a spectrum of previously undiagnosed low γ‐glutamyltransferase cholestasis. Hepatology (Baltimore, Md.), 65(5), 1655-1669.
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9

Histological and Immunohistochemical Analysis of Liver Tissue

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Liver tissue was formalin-fixed and paraffin-embedded. Hematoxylin-eosin (HE), periodic acid-Schiff (PAS), PAS after diastasis, and trichrome of Masson were routinely analyzed. Immunohistochemistry was made with anti-cytokeratin7 (CK7) (monoclonal mouse anti-human, OV-TL12/30, ready to use; Agilent, Santa Clara, CA, USA), anti-CK19 (monoclonal mouse anti-human, RCK108, ready to use; Agilent), anti-MYO5B (rabbit polyclonal, C-term human MYO5B, LS-B3118/122815, Life Span BioSciences, Inc, Seattle, WA, USA, 1:100) and anti-BSEP (rabbit polyclonal, NBP1-89319, Novus Biologicals, Centennial, CO, USA, 1:1000). Normal liver (donor's livers at time 0) was used as a positive control.
Bioinformatics Analysis and Ultrastructural analysis: see supplemental content, http://links.lww.com/MPG/C679.
Matarazzo L., Bianco A.M., Athanasakis E., Serveres M., Francalanci P., Cenacchi G., Maggiore G, & D'Adamo A.P. (2022). MYO5B Gene Mutations: A Not Negligible Cause of Intrahepatic Cholestasis of Infancy With Normal Gamma-Glutamyl Transferase Phenotype. Journal of pediatric gastroenterology and nutrition, 74(5).
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