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Bond 3 system

Manufactured by Leica
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The Bond-III system is a piece of laboratory equipment manufactured by Leica. It is designed to perform automated sample preparation and analysis. The core function of the Bond-III system is to enable researchers and scientists to process and analyze samples efficiently and consistently.

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Lab products found in correlation

Bond polymer refine detection kit, by Leica (5 mentions) Refine hrp kit, by Leica (3 mentions) Bond polymer refine detection, by Leica (2 mentions) Ar9640, by Leica (2 mentions) Hpa003711, by Merck Group (2 mentions) Bond epitope retrieval 2, by Cell Signaling Technology (2 mentions) Bond polymer kit, by Cell Signaling Technology (2 mentions) E1l3n, by Cell Signaling Technology (2 mentions) Vectastain elite abc kit, by Vector Laboratories (2 mentions) Biotin sp conjugated donkey anti rabbit igg, by Jackson ImmunoResearch (2 mentions) Hhv 8 orf73, by Leica (2 mentions) Clone 2a3, by Fortis Life Sciences (2 mentions) Bl 95 1a2, by Fortis Life Sciences (2 mentions) Dm2500 microscope, by Leica (1 mentions) Application suite version 2.8.1, by Leica (1 mentions) Ab92309, by Abcam (1 mentions) Ap1802a, by Abcepta (1 mentions) Bond epitope retrieval solution 2, by Leica (1 mentions) Bond epitope retrieval solution 1, by Leica (1 mentions) Anti pecam1 cd31, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Bond III (387 citations) BOND III automated system (8 citations) BOND‐MAX and BOND‐III systems (2 citations) Leica BOND-III staining system (2 citations) BOND-III automated immunostaining system (2 citations) Bond-III™ Polymer Refine Detection system (2 citations)

34 protocols using bond 3 system

1

Immunohistochemical Analysis of Ikaros Expression

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50–250 thousand cells were centrifuged onto glass slides and fixed for 20 minutes in 10% neutral buffered formalin. Immunohistochemical stains were performed on a Bond III system (Leica Microsystems, Bannockburn, Ill) with pH6 epitope retrieval solution (Leica), a HRP-conjugated anti-Ikaros primary antibody (ab26083, Abcam, Cambridge, MA) diluted 1:1000 in IHC diluent (Leica), and with nuclear counter stain hematoxylin, following manufacturer’s protocol (standard protocol F, Leica) but eliminating steps to de-paraffinize slides. Stained slides were analyzed on a Leica DM 2500 microscope with a 40x HCX PL Fluotar objective (∞/0.17/D). Images were captured using Leica application suite version 2.8.1 (build 1554, 2003–2007).
O’Brien S., Thomas R.M., Wertheim G.B., Zhang F., Shen H, & Wells A.D. (2014). Ikaros imposes a barrier to CD8+ T cell differentiation by restricting autocrine IL-2 production. Journal of immunology (Baltimore, Md. : 1950), 192(11), 5118-5129.
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2

Automated Immunohistochemical Staining of FFPE Tissues

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After the tumor center and invasive margin area were reviewed, pathologic slides from FFPE tissues were stained with monoclonal antibodies to CD3 and CD8. Immunostaining for CD3 and CD8 (Leica Microsystems, Newcastle-upon-Tyne, UK) was performed on the same completely automated Bond-III system (Leica Microsystems) using onboard heat-induced antigen retrieval with epitope retrieval solution 2 for 10 min at 99°C, followed by incubation with the antibody for 30 min at room temperature. This automated system used a Refine polymer detection kit with horseradish peroxidase polymer as a secondary antibody and DAB, and incubation with secondary antibody was performed for 30 min at room temperature.
Ishii H., Azuma K., Kawahara A., Matsuo N., Tokito T., Kinoshita T., Yamada K., Sasada T., Akiba J, & Hoshino T. (2017). Programmed cell death-ligand 1 expression and immunoscore in stage II and III non-small cell lung cancer patients receiving adjuvant chemotherapy. Oncotarget, 8(37), 61618-61625.
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3

Evaluating BAG3 and EGFR in TNBC Prognosis

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Full tissue sections were obtained from a cohort of 80 patients (Cohort 3) diagnosed with Triple Negative Breast Cancer at St. Vincent’s University Hospital, Dublin Ireland between 2000 and 2010 who had not received chemotherapy. All the patients provided an informed consent for using their samples. In this study, patients were censored at 10 years with a median age of 67 years (range between 32–99 years).
Deparaffinisation, antigen retrieval and IHC staining for BAG3 and EGFR were performed on an automated platform (Bond™ III system – Leica MicroSystems™, Newcastle, U.K.). Staining for BAG3 was performed using a rabbit monoclonal antibody (anti- BAG3 Abcam cat# ab92309; 1:300 dilution, ER1 antigen retrieval for 20 minutes). For EGFR immunostaining, a mouse monoclonal antibody (Leica Biosystems NCL-L-EGFR-384; 1:100 dilution, ER1 antigen retrieval for 20 minutes) was used. Antibody optimisation was performed on invasive breast cancer (BAG3) and placental (EGFR) TMAs.
Histopathological review of the sections was performed for BAG3 and the tissues were graded initially as (0, 1, 2, 3) and further graded as either high BAG3 expressing tissues (Score 2, 3) or as low BAG3 (Score 0, 1) expressing tissues. Survival curves based on Kaplan-Meier estimates were used to determine the relationship between BAG3 and recurrence free survival in this Cohort (Cohort 3).
Shields S., Conroy E., O’Grady T., McGoldrick A., Connor K., Ward M.P., Useckaite Z., Dempsey E., Reilly R., Fan Y., Chubb A., Matallanas D.G., Kay E.W., O’Connor D., McCann A., Gallagher W.M, & Coppinger J.A. (2018). BAG3 promotes tumour cell proliferation by regulating EGFR signal transduction pathways in triple negative breast cancer. Oncotarget, 9(21), 15673-15690.
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4

Automated Immunohistochemical Staining

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All the tissue slides were stained by the fully automated Bond-III system (Leica Microsystems, Newcastle-upon-Tyne, UK) according to the manufacturer’s instructions. The following primary antibodies were used: PD-L1 (MXR003; 1:750; MXB Biotechnologies, Fuzhou, China) and CD8 (clone NCL-L-CD8-4B11; 1: 100; DAKO, Minneapolis, MN, USA).
Wang W., Jing H., Liu J., Bu D., Zhang Y., Zhu T., Lu K., Xu Y., Cheng M., Liu J., Yao J., Huang S, & Wang L. (2021). Correlation between schistosomiasis and CD8+ T cell and stromal PD-L1 as well as the different prognostic role of CD8+ T cell and PD-L1 in schistosomal-associated colorectal cancer and non-schistosomal-associated colorectal cancer. World Journal of Surgical Oncology, 19, 321.
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5

Automated Immunohistochemistry for LC3B and Cleaved Caspase-3

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Deparaffinization, antigen retrieval and IHC staining for LC3B and cleaved caspase-3 were performed on an automated platform (Bond™ III system, Leica MicroSystems™, Milton Keynes, U.K.). For LC3B (Atg8b) immunostaining, a rabbit polyclonal antibody was used (ABGENT cat# AP1802a) at 1:100 dilution. Antigen retrieval is an automated process on the Bond instrument and involves the tissue being heated for 10 minutes at 100°C in ER1 retrieval buffer (BondTM Epitope Retrieval, Leica MicroSystems™, Milton Keynes, U.K.). Cleaved caspase-3 immunostaining was performed using a rabbit monoclonal antibody (Cell Signaling Technology cat# 9664; 1:200 dilution, ER1 antigen retrieval for 10 minutes). Primary antibody binding was visualized using the Bond™ Polymer Refine Detection containing a peroxide block, post primary, polymer reagent, DAB chromogen and haematoxylin counterstain.
El Mashed S., O’Donovan T.R., Kay E., O’Grady A., McManus D., Turkington R.C, & McKenna S.L. (2022). Apoptosis and autophagy markers predict survival in neoadjuvant treated oesophageal adenocarcinoma patients. BMC Cancer, 22, 908.
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6

MHC-II Immunohistochemical Staining Protocol

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Immunohistochemical staining for MHC-II was performed by the MSKCC Pathology Department using purified mouse antibody clone LGII-612.14 provided by Dr. Soldano Ferrone, (Massachusetts General Hospital, Boston, MA) at 1:200 dilution (0.5 μg/mL). Staining was performed on the Leica Bond-III system (Leica, Buffalo Grove, IL), using heat-based antigen retrieval, a high pH buffer solution (AR9640; Leica, Bond Epitope Retrieval Solution 2, 30 minutes), 30-minute primary incubation time, and a polymer detection system (DS9800; Leica, Bond Polymer Refine Detection).
Redelman-Sidi G., Binyamin A., Antonelli A.C., Catalano W., Bean J., Al-Ahmadie H., Jungbluth A.A, & Glickman M.S. (2022). BCG-induced tumor immunity requires tumor-intrinsic CIITA independent of MHC-II. Cancer immunology research, 10(10), 1241-1253.
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7

SSTR2 Immunohistochemistry Protocol

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Chromogenic in situ hybridization for HPV and EBER, and IHC for p40 and p16 have been described previously (17 (link)–19 (link)). For SSTR2, IHC was performed using SSTR2 antibody (1:800, Rabbit monoclonal antibody, Clone UMB-7, Abcam, City, State) on formalin fixed paraffin-embedded tissue sections on a Leica Bond III system (Buffalo Grove, IL). The section was pre-treated using heat mediated antigen retrieval with Bond ER1 antigen retrieval buffer for 30 minutes and incubated with the antibodies for 15 min at room temperature. SSTR2 was detected with Leica Bond Refine detection system, and each section was counterstained with hematoxylin. The extent of staining is categorized as focal, multifocal, or diffuse, and the intensity of the staining is categorized as weak, moderate or strong. Staining of rare tumor cells (<1%) was considered negative. Focal to diffuse staining of any intensity was considered positive and the number of positive cases in each tumor subset was determined.
Viswanathan K, & Sadow P.M. (2021). Somatostatin receptor 2 (SSTR2) is highly sensitive and specific for Epstein-Barr Virus-associated nasopharyngeal carcinoma. Human pathology, 117, 88-100.
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8

Immunohistochemical Analysis of NPNT and CYP11B2

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Immunohistochemistry was carried out using the peroxidase-antiperoxidase method on fresh frozen human tissue. In cases where fresh frozen tissue was unavailable, immunohistochemistry was performed on formalin-fixed, paraffin-embedded adrenal sections (4 μm) using an automated immunostainer with cover tile technology (Bond-III system, Leica Biosystems). NPNT antibody (HPA003711, Sigma; 1:50 dilution), and CYP11B2 antibody (custom mouse anti-human antibody from Dr Celso E. Gomez-Sanchez)22 (link) were used as the primary antibodies. Further details are provided in the Data Supplement.
Teo A.E., Garg S., Johnson T.I., Zhao W., Zhou J., Gomez-Sanchez C.E., Gurnell M, & Brown M.J. (2017). Physiological and Pathological Roles in Human Adrenal of the Glomeruli-Defining Matrix Protein NPNT (Nephronectin). Hypertension (Dallas, Tex. : 1979), 69(6), 1207-1216.
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9

Immunohistochemical Evaluation of MYC and BCL6

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MYC (Abcam clone Y69) and BCL6 (Leica Clone LN22) immunohistochemistry were performed where possible in all cases where tissue materials remained available using the Bond-III system (Leica Biosystems) with the Bond Polymer Refine Detection Kit as the same condition of routine histopathological diagnosis. This was carried out centrally in the Cambridge lab and the staining intensity (weak, moderate, strong) and percentage in tumour cells (>70% or <70%) were scored [11 (link)].
Zhang C., Stelloo E., Barrans S., Cucco F., Jiang D., Tzioni M.M., Chen Z., Li Y., Swennenhuis J.F., Makker J., Rásó-Barnett L., Liu H., El-Daly H., Soilleux E., Shah N., Nagumantry S.K., Kyaw M., Prahladan M.P., Tooze R., Westhead D.R., Feitsma H., Davies A.J., Burton C., Johnson P.W, & Du M.Q. (2024). Non-IG::MYC in diffuse large B-cell lymphoma confers variable genomic configurations and MYC transactivation potential. Leukemia, 38(3), 621-629.
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10

Automated PD-L1 Immunohistochemistry in FFPE Tissue

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This was performed on FFPE tissue sections using an automated immunostainer (Bond-III system, Leica Biosystems). Following antigen retrieval by combination of heat and Bond Epitope Retrieval 2 solution for 20 minutes, PD-L1 was stained with the monoclonal antibody clone E1L3N (Cell Signalling) and visualized using Bond Polymer Kit.
Wu F., Watanabe N., Tzioni M.M., Akarca A., Zhang C., Li Y., Chen Z., Cucco F., Carmell N., Noh J.Y., Ito K., Dobson R., Moody S., Yao W., Zhang W., Liu W., Liu H., Okosun J., Chott A., Bi Y., Chuang S.S., Raderer M., Li J.Y., Marafioti T, & Du M.Q. (2021). Thyroid MALT lymphoma: self-harm to gain potential T-cell help. Leukemia, 35(12), 3497-3508.
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