Bond polymer detection kit

Manufactured by Leica Biosystems

The Bond Polymer detection kit is a laboratory equipment product designed for use in immunohistochemistry (IHC) procedures. It provides a polymer-based detection system for the visualization of target antigens in tissue samples. The kit includes the necessary reagents and components to perform the staining process, enabling researchers and pathologists to detect the presence and localization of specific proteins or molecules of interest within biological specimens.

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Lab products found in correlation

Autostainer xl, by Leica camera (1 mentions) Ventana benchmark xt staining system, by Roche (1 mentions) Leica bond max autostainer, by Leica Biosystems (1 mentions) Ventana ultra view universal dab detection kit, by Roche (1 mentions) Cytoseal xyl, by Thermo Fisher Scientific (1 mentions) Leica bond 3 autostainer, by Leica Biosystems (1 mentions) Aperio at2 scanner, by Leica Biosystems (1 mentions)

3 protocols using bond polymer detection kit

PanIN Lesion Validation and Multiplex Imaging

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When PanIN lesions were detected in a 350 µm tissue section via 3D imaging, the section was processed by dehydration, embedding, and microtome sectioning to generate 4 µm paraffin slices for H&E staining (Leica Autostainer XL). H&E‐stained specimens were examined using the same Zeiss microscope for a side‐by‐side comparison of the 3D fluorescence and 2D H&E micrographs to confirm the PanIN lesion. Once confirmed, multiplex signals from the same microenvironment were further acquired from adjacent sections via standard immunohistochemistry (4 µm sections; Bond Polymer detection kit, Leica Biosystems #DS9800/DS9390) and 3D histology (350 µm sections) as described previously.^[⁴⁵ (link)
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Hu C., Huang C., Hsu M., Chien H., Wu P., Chen Y., Jeng Y., Tang S., Chung M., Shen C., Chang M., Chang Y., Tien Y, & Lee W. (2023). Oncogenic KRAS, Mucin 4, and Activin A‐Mediated Fibroblast Activation Cooperate for PanIN Initiation. Advanced Science, 10(36), 2301240.

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Optimization of Tissue Immunohistochemistry

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Four-µm-thick tissue sections on glass slides were deparaffinized at 60 °C for 1 hour, dewaxed at 72 °C for 3 minutes, and hydrated by washing with alcohol three times. Immunohistochemical staining was performed using two types of autostainer, Leica Bond-max autostainer (Leica Biosystems, Melbourne, Australia) and Ventana BenchMark XT staining system (Ventana Medical Systems, Inc., Tucson, AZ, USA). Antigens were retrieved by heat treatment in the epitope retrieval solution for each autostainer (pH 6.0 Bond epitope retrieval solution 1, pH 9.0 Bond epitope retrieval solution 2, or Ventana mild 32 condition reagent). After peroxidase blocking for 5 minutes, the primary antibody was incubated for 15 minutes. The type, dilution level, and company information of the 25 primary antibodies used are listed in Table 5. After the primary antibody reaction, a chromogenic procedure was done using the Bond polymer detection kit (Leica Biosystems, Cat. No. DS9800) or Ventana ultraView universal DAB detection kit (Ventana Medical Systems, Cat. No. 760-500). Counterstaining for nuclei was done by incubation with hematoxylin for 1 minute.

Lee K.B., Lee K.S, & Lee H.S. (2019). Tumor-Associated Protein Profiles in Kaposi Sarcoma and Mimicking Vascular Tumors, and Their Pathological Implications. International Journal of Molecular Sciences, 20(13), 3142.

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Immunohistochemistry Analysis of FFPE Samples

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Formalin-fixed paraffin-embedded (FFPE) cell lines/tissue of 4-μm thickness slides were used to perform all IHCs using Leica Bond III Auto Stainer (Leica Biosystems) except HER2 that was performed using Ventana Benchmark ULTRA IHC/ISH platform (Roche). In brief, after deparaffinization with bond dewax (cat no. AR9222; Leica Biosystems) solution followed by heat-mediated epitope retrieval [Bond TM Epitope Retrieval Solution 1&2 (Leica Biosystem) cat nos. AR9961 and AR9640, respectively] were blocked by peroxidase blocking (3% H₂O₂) for 20 minutes. After blocking, slides were incubated for 15 minutes with the following primary antibodies. All the antibodies were detected using a Bond polymer Detection kit (cat no. DS9800, Leica Biosystem) with diaminobenzidine (DAB) as a chromogen. Supplementary Table S4 provides detailed antibody information. Subsequent transient washing, postprimary, and polymer incubation, followed by hematoxylin staining, were performed. Finally, slides were air dried, and cover slipped with mounting media (Cytoseal XYL, Thermo Scientific). and digitalized in an Aperio AT2 scanner (Leica Biosystems; Vista) under a 20× objective magnification.

DiPeri T.P., Evans K.W., Wang B., Zhao M., Akcakanat A., Raso M.G., Rizvi Y.Q., Zheng X., Korkut A., Varadarajan K., Uzunparmak B., Dumbrava E.E., Pant S., Ajani J.A., Pohlmann P.R., Jensen V.B., Javle M., Rodon J, & Meric-Bernstam F. (2024). Co-clinical Trial of Novel Bispecific Anti-HER2 Antibody Zanidatamab in Patient-Derived Xenografts. Cancer Discovery, 14(5), 828-845.

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