Bond max automated ihc ish stainer

Manufactured by Leica

Sourced in Germany

The BOND-MAX Automated IHC/ISH Stainer is a laboratory instrument designed for performing immunohistochemistry (IHC) and in situ hybridization (ISH) staining procedures. It automates the entire staining process, including reagent application, incubation, and washing steps, to ensure consistent and reliable results.

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Lab products found in correlation

Bond polymer refine detection kit, by Leica (2 mentions) Ab22510, by Abcam (1 mentions) Ab56417, by Abcam (1 mentions) Ab68672, by Abcam (1 mentions) Ab216327, by Abcam (1 mentions) Ncl l cd4 368, by Leica (1 mentions) Ncl l cd8 4b11, by Leica (1 mentions) Sc 100289, by Santa Cruz Biotechnology (1 mentions) Mca1477, by Bio-Rad (1 mentions) Dm2000 led microscope, by Leica (1 mentions) Dfc7000 t camera, by Leica (1 mentions) Ab51603, by Abcam (1 mentions) Ab112, by Beyotime (1 mentions)

Close spelling variants of this product

Bond-Max Automated IHC stainer Bond Max IHC stainer BOND automated IHC Stainer Leica BOND-MAX stainer Automated stainer Bond Max Leica stainer

7 protocols using bond max automated ihc ish stainer

Comprehensive Immunohistochemical Profiling of Tissues

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Paraffin blocks were cut into 1 µm-thick sections and fixed onto a poly-L-lysine treated glass slide and then used for histological and immunohistochemical analyses. All sections were stained with H&E following manufacturer’s protocol to achieve correct orientation of the biopsy. Immunohistochemical staining was performed using Bond Polymer refine Detection kit on a BOND-MAX Automated IHC/ISH Stainer (Leica Biosystems) following the manufacturer’s protocol. The following antibodies were used: mouse monoclonal anti-human CD19 (NCL-L-CD19-163, Leica Biosystems), rat anti-human CD3 (MCA1477,Bio-Rad), mouse monoclonal anti-human CD4 (NCL-L-CD4-368, Leica Biosystems), mouse monoclonal anti-human CD8 (NCL-L-CD8-4B11, Leica Biosystems), mouse monoclonal anti-human Claudin-1 (ab56417, Abcam), mouse monoclonal anti-human E-Cadherin (36B5) (PA0387, Leica Biosystems), mouse monoclonal anti-human FoxP3 (ab22510, Abcam), mouse monoclonal anti-human γδ-TCR (sc-100289, Santa Cruz Biotechnology), mouse monoclonal anti-human Langerin (ab49730, Abcam), rabbit recombinant monoclonal [EPR20992] to Occludin (ab216327, Abcam), and rabbit polyclonal to Neutrophil Elastase (ab68672, Abcam). Positive and negative controls were included for each experiment.

Sanchez-Solares J., Sanchez L., Pablo-Torres C., Diaz-Fernandez C., Sørensen P., Barber D, & Gomez-Casado C. (2021). Celiac Disease Causes Epithelial Disruption and Regulatory T Cell Recruitment in the Oral Mucosa. Frontiers in Immunology, 12, 623805.

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Quantification of eIF4E Phosphorylation in Mouse Brains

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After sacrificing mice, resected brains were harvested for hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) analysis. Brains were fixed with 10% buffered formalin overnight. Brains were then embedded in paraffin and sectioned for H&E staining and IHC analysis. Sections were processed using the BOND-MAX Automated IHC/ISH Stainer and its Polymer Detection System (Leica Biosystems). The Bond Dewax Solution (AR9222) was used at 72°C. The Bond Epitope Retrieval Solution 1 (AR9961) was used for 20 min at 100°C. Samples were pretreated with 3% hydrogen peroxide for 5 minutes. For detection of eIF4E phosphorylation, the eIF4E (Ser209) antibody [EP2151Y] (Abcam) was used at a 1:2000 dilution for 15 minutes. The post primary polymer penetration enhancer reagent was added for 8 minutes followed by the polymer poly-HRP secondary antibody for 8 minutes. Hematoxylin was used for 5 minutes. eIF4E phosphorylation (Ser209) was semiquantified by light microscopic analysis, in which a board-certified neuropathologist (CH) ranked the tumors from strongest to weakest, followed by Spearman rank order correlation. Mitoses were scored per 10 high power fields (600x). Both phospho-eIF4E and mitotic index were analyzed while blinded to treatment group.

Bell J.B., Eckerdt F., Alley K., Magnusson L.P., Hussain H., Bi Y., Arslan A.D., Clymer J., Alvarez A.A., Goldman S., Cheng S.Y., Nakano I., Horbinski C., Davuluri R.V., James C.D, & Platanias L.C. (2016). MNK Inhibition Disrupts Mesenchymal Glioma Stem Cells and Prolongs Survival in a Mouse Model of Glioblastoma. Molecular cancer research : MCR, 14(10), 984-993.

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Immunohistochemical Staining of B7-H6 in Cervical Biopsies

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Using the paraffin-embedded cervical biopsies, 2 μm thick sections were cut. Sections were de-waxed with xylene and rehydrated with ethanol and finally water. Antigen retrieval was performed using citrate buffer pH 6, 95 °C for 25 min. Hydrogen peroxide (3–4%) was applied to block endogenous peroxidase and then rinsed with TBS; after washing, sections were incubated with primary rabbit anti-human B7-H6 polyclonal antibody (Abcam, Cambridge, MA, catalog number 121794), diluted 1:250 in antibody diluent from Leica for 35 min. Sections were rinsed with TBS and then incubated with post-primary antibody solution for 10 min, then again rinsed and horseradish peroxidase-labeled IgG anti-rabbit was applied for 15 min. Then, a solution containing 66 mM 3,3′-diaminobenzidine tetrahydrochloride hydrate and other stabilizer solutions containing (≤0.1%) hydrogen peroxide were applied for 3 min; subsequently, sections were rinsed with water and stained with hematoxylin < 0.1% 4 min. All steps were performed using the BOND-MAX Automated IHC/ISH Stainer (Leica Biosystems).

Gutierrez-Silerio G.Y., Franco-Topete R.A., Haramati J., Navarrete-Medina E.M., Gutierrez-Franco J., Bueno-Topete M.R., Bastidas-Ramirez B.E., Ramos-Marquez M.E, & del Toro-Arreola S. (2020). Positive staining of the immunoligand B7-H6 in abnormal/transformed keratinocytes consistently accompanies the progression of cervical cancer. BMC Immunology, 21, 9.

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Tissue Microarray Construction and Immunohistochemistry

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Three representative foci in each case were selected for TMA blocks. Three cores (2 mm in diameter) were obtained from each paraffin block to make a TMA block. Serial sections were prepared and stained with hematoxylin and eosin. Immunohistochemistry was carried out on the TMA slides using a Bond-Max automated IHC/ISH stainer (Leica Biosystems, Wetzlar, Germany) or a Ventana system (Benchmark Ultra, Tucson, Arizona, USA) with primary antibodies against the following proteins:

Kim S.S., Cho Y.M., Kim G.H., Kee K.H., Kim H.S., Kim K.M., Kim J.H, & Choi C. (2020). Recurrent KRAS mutations identified in papillary renal neoplasm with reverse polarity-a comparative study with papillary renal cell carcinoma. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 33(4).

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Immunohistochemical Analysis of Neurodegenerative Markers

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Histological examination was performed on 5 μm thick sections cut from formalin-fixed, paraffin-embedded tissue of frontal, pre- and post-central, and temporal cortex, hippocampus and spinal cord. Sections were stained with antibodies against pTDP-43, TUBA4A (C-term), pTau^(S202/T205) and Aβ_17–24 (Online resource Supplementary Table 2). Stainings were performed with the BOND-MAX automated IHC/ISH Stainer (Leica Biosystems, Wetzlar, Germany) using the Bond Polymer Refine Detection kit (DS9800, Leica Biosystems). Briefly, slides were deparaffinized and epitopes were retrieved with low or high pH buffer. After incubation with Peroxidase-Blocking Reagent (DS9800, Leica Biosystems), slides were incubated with primary antibodies for 30 min, followed by secondary antibody incubation. DAB was used for visualization, followed by counterstaining with hematoxylin. Dehydration was carried out in an autostainer, followed by mounting in an automated cover-slipper (Leica Biosystems). Images were acquired using the Leica DM2000 LED microscope coupled to a Leica DFC 7000 T camera. Images were processed using ImageJ and combined into figures using Inkscape.

Van Schoor E., Strubbe D., Braems E., Weishaupt J., Ludolph A.C., Van Damme P., Thal D.R., Bercier V, & Van Den Bosch L. (2024). TUBA4A downregulation as observed in ALS post-mortem motor cortex causes ALS-related abnormalities in zebrafish. Frontiers in Cellular Neuroscience, 18, 1340240.

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Immunohistochemical Analysis of DYNLL1 and BCL2

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Immunohistochemical staining of DYNLL1 and BCL2 was performed using primary antibodies against DYNLL1 (ab51603, 1:600; Abcam) and BCL2 (AB112, 1:200; Beyotime Biotechnology) on tissue microarray sections by a BOND-MAX Automated IHC/ISH Stainer (Leica). The immunostaining degree of the DYNLL1 and BCL2 proteins was evaluated as previously described [20 (link)] by pathologists based on the nuclear staining intensity (intensity score) and percentage of positive cells (extent score). The final immunoreactivity score for each sample was the product of the intensity score and extent score.

Li Q., Zhang Z., Jiang H., Hou J., Chai Y., Nan H., Li F, & Wang L. (2022). DLEU1 promotes cell survival by preventing DYNLL1 degradation in esophageal squamous cell carcinoma. Journal of Translational Medicine, 20, 245.

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Immunohistochemical Staining of SOX2 and YAP1

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Immunohistochemical stainings of SOX2 and YAP1 were performed using primary antibodies against SOX2 (#2748, Cell Signaling Technology) and YAP1 (#14074, Cell Signaling Technology) on a BOND‐MAX Automated IHC/ISH Stainer (Leica) according to previously established protocols.24 Following staining, tissue microarray sections were dehydrated in graded alcohol, cleared in xylene, and mounted.
Immunostaining degree of each sample was scored as previously described by pathologists based on nuclear staining intensity (intensity score) and percentage of positive cells (extent score).6 The final immunoreactivity score for each case is the product of the intensity score and the extent score.

Chai Y., Li Q., Zhao H., Zhang Z., Yu X., Pang L., Liu Z., Zhao J., Wang L, & Li F. (2019). SOX2 antagonizes WWC1 to drive YAP1 activation in esophageal squamous cell carcinoma. Cancer Medicine, 8(16), 7055-7064.

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