Bond polymer refine kit

Manufactured by Leica camera

The Bond Polymer Refine kit is a laboratory equipment product designed for specimen preparation. It is used to process and refine samples for analysis. The kit contains the necessary components to perform this process, though the specific technical details and intended use are not available in this factual and unbiased format.

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Lab products found in correlation

Anti hmgb1, by Abcam (2 mentions) Anti cd3, by Abcam (2 mentions) Rabbit anti cd68 antibody, by Abcam (1 mentions) Bond polymer refine kit, by Leica Biosystems (1 mentions) Nanozoomer digital pathology system, by Hamamatsu Photonics (1 mentions) Bond max autostainer, by Leica camera (1 mentions) Ndp system, by Hamamatsu Photonics (1 mentions) Concentrate, by Leica camera (1 mentions) Vectra 3.0 automated quantitative pathology imaging system, by Akoya Biosciences (1 mentions) Bond rx system, by Leica camera (1 mentions) Bond polymer refine red kit, by Leica camera (1 mentions) Halo image analysis platform, by Indica Labs (1 mentions) Ab32072, by Abcam (1 mentions)

7 protocols using bond polymer refine kit

Immunostaining of Formalin-Fixed Liver

Cited 1 time

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Formalin-fixed paraffin-embedded liver sections were used for immunohistochemistry which was performed in a similar manner as reported earlier [31 (link)]. Overnight incubation at 4 °C was performed using anti-HMGB1 (Abcam), anti-IBA-1 (Waco), and anti-CD-3 (Abcam) primary antibodies. Secondary detection was performed via Leica Bond Polymer Refine kit and slides were counterstained with hematoxylin.

Bulant M., Roussel J.P., Astier H., Nicolas P, & Vaudry H. (1990). Processing of thyrotropin-releasing hormone prohormone (pro-TRH) generates a biologically active peptide, prepro-TRH-(160-169), which regulates TRH-induced thyrotropin secretion.. Proceedings of the National Academy of Sciences of the United States of America, 87(12), 4439-4443.

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Quantifying Tissue Pathology Markers

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All immunohistochemistry staining was performed in 5-μm paraffin sections with BondRX Stainer. Briefly, the primary antibodies including rabbit anti-mouse IgG from Bond Polymer Refine kit (Leica Biosystems, Buffalo Grove, IL, ready to use), rabbit anti-CD68 antibody (Abcam, ab125212, 1:500), and rabbit anti-Collagen1 antibody (BosterBio, Pleasanton, CA, PA2140-2, 1:1000) were used for detection of necrosis, inflammation, and fibrosis, respectively. Bond Polymer Refine kit (Leica, Cat No: DS9800) was applied as the detection system. The positive cells were identified as brown in color and nuclei were stained blue. The stained slides were scanned with Aperio ScanScope AT2 scanner. The whole digital slides were viewed and analyzed by ImageScope. The positive pixel count algorithm was selected and adjusted to cover each individual positive staining for analysis. The data was presented as positivity which was obtained from the following formula: positivity (%) = positive area (pixels)/total analyzed area (pixels) × 100%.

Iskenderian A., Liu N., Deng Q., Huang Y., Shen C., Palmieri K., Crooker R., Lundberg D., Kastrapeli N., Pescatore B., Romashko A., Dumas J., Comeau R., Norton A., Pan J., Rong H., Derakhchan K, & Ehmann D.E. (2018). Myostatin and activin blockade by engineered follistatin results in hypertrophy and improves dystrophic pathology in mdx mouse more than myostatin blockade alone. Skeletal Muscle, 8, 34.

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Immunohistochemical Analysis of Liver Oxidative Stress

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Immunohistochemistry was performed on paraffin embedded liver sections. Briefly, the slides were first deparaffinized using xylene and dehydrated using ethanol. Theses slides were then pressure heated at 100 °C for 20 min in Na-citrate buffer. To inactivate the endogenous peroxide activity slides were kept for 12 min at room temperature in 3% H₂O₂ in TBS. Further, the slides were incubated with blocking buffer for 30 min to block non-specific binding sites followed by overnight incubation in anti 4-HNE primary antibody (Abcam). Detection was performed using Leica Bond Polymer Refine kit and slides were counterstained with hematoxylin. The stained slides were scanned using Hamamatsu Nanozoomer Digital Pathology system (Hamamatsu City, Japan) and the digital information were stored for analysis.

Kumar A., Pathak R., Palfrey H.A., Stone K.P., Gettys T.W, & Murthy S.N. (2020). High levels of dietary methionine improves sitagliptin-induced hepatotoxicity by attenuating oxidative stress in hypercholesterolemic rats. Nutrition & Metabolism, 17, 2.

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Immunostaining Protocol for ER and Aurora A

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Immunostaining was performed using a Bond polymer refine kit and a Leica Bond-max autostainer (Supplementary Table 8). Immunohistochemical data for ER and aurora kinase A expression have been previously described in detail.^{34 (link), 38 (link), 39 (link)} Negative controls were performed simultaneously. Assay specificity was checked using cytoblocks prepared from cultured cells with siRNA knockdown (Supplementary Figure 2). Nine from a total of 118 arrays stained were deemed technical failures and excluded.

Modelska A., Turro E., Russell R., Beaton J., Sbarrato T., Spriggs K., Miller J., Gräf S., Provenzano E., Blows F., Pharoah P., Caldas C, & Le Quesne J. (2015). The malignant phenotype in breast cancer is driven by eIF4A1-mediated changes in the translational landscape. Cell Death & Disease, 6(1), e1603-.

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Immunohistochemical Analysis of Liver Tissue

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For immunohistochemistry, formalin-fixed paraffin-embedded liver sections were used. Slides were briefly deparaffinized with xylene, dehydrated with ethanol and then pressure heated for 20 min at 100 °C in Na citrate buffer. Endogenous peroxidase activities were inactivated in 3% H₂O₂ in TBS for 12 min at room temperature. Samples were then incubated with protein block for 30 min to block non-specific antibody-binding sites. Overnight incubation at 4 °C was performed using anti-IBA-1 (Waco), anti-CD-3 (Abcam) and anti-HMGB1 (Abcam) primary antibodies. A negative antibody control was included in each case by replacing the primary antibody with antibody diluent. Secondary detection was performed via Leica Bond Polymer Refine kit and slides were counterstained with hematoxylin. Slides were scanned using a Hamamatsu NDP system (Hamamatsu City, Japan). Using 4–6 random fields per slide, positive staining was quantified and analyzed with the help of ImageJ software.

Pathak R., Kumar A., Palfrey H.A., Forney L.A., Stone K.P., Raju N.R., Gettys T.W, & Murthy S.N. (2019). The incretin enhancer, sitagliptin, exacerbates expression of hepatic inflammatory markers in rats fed a high-cholesterol diet. Inflammation research : official journal of the European Histamine Research Society ... [et al.], 68(7), 581-595.

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RNAscope LS2.5 In-Situ Hybridization

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Following sample pretreatment, hybridization and amplification steps were done according to the RNAscope LS2.5 protocol (ACD, UM-322150 RevA). Probes were hybridized for 2 hours at 42°C. Slides were washed with 1X Bond wash buffer (Leica 10X concentrate, AR9590) at 42ºC 3 times (0, 1, 5 minutes) followed by 8 washes with 1X Bond wash buffer 0 minute each. Slides were then treated with Amp 1 to Amp 6 steps using RNAscope 2.5 LS Reagents Kit-Brown (ACD, 322100).
ISH detection and counterstain were completed using Bond Polymer Refine kit (Leica, DS9800).

Chan S., Filézac de L’Etang A., Rangell L., Caplazi P., Lowe J.B, & Romeo V. (2018). A method for manual and automated multiplex RNAscope in situ hybridization and immunocytochemistry on cytospin samples. PLoS ONE, 13(11), e0207619.

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Multiplex IHC for Cell Cycle Markers

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Dual IHC staining of Ki67 (Biocare Medical, CRM325, RRID: AB_2721189), ER (Thermo Fisher Scientific RM9101S0, RRID: AB_149902), p-ER (S118; SAB 11072, RRID: AB_895302), p-CDK1 (T161; Cell Signaling Technology, CST9114, RRID: AB_2074652), p-CDK2 (T160; Cell Signaling Technology, CST2561, RRID: AB_2078685), and c-Myc (Abcam, ab32072, RRID: AB_731658) was conducted on 4-μm FFPE sections, using both Bond Polymer Refine Kit and Bond Polymer Refine Red kit in Leica Bond RX system. The slides were deparaffinized and heat-mediated antigen retrieval was performed with EDTA buffer (pH 9.0). The IHC staining was performed using the antibodies and the incubation conditions reported in Supplementary Table S8. Antigen-antibody reaction was visualized with 3,3′-diaminobenzidine (DAB) chromogen. Omission of the primary antibody was used as a negative control. Whole slide images were acquired from stained slides using a Vectra 3.0 Automated Quantitative Pathology Imaging System (Akoya Biosciences) and analyzed using Halo Image Analysis platform (Indica Labs). Image annotations were performed by one research pathologist. Areas containing invasive carcinoma were included in image analysis.

Guarducci C., Nardone A., Russo D., Nagy Z., Heraud C., Grinshpun A., Zhang Q., Freelander A., Leventhal M.J., Feit A., Cohen Feit G., Feiglin A., Liu W., Hermida-Prado F., Kesten N., Ma W., De Angelis C., Morlando A., O'Donnell M., Naumenko S., Huang S., Nguyen Q.D., Huang Y., Malorni L., Bergholz J.S., Zhao J.J., Fraenkel E., Lim E., Schiff R., Shapiro G.I, & Jeselsohn R. (2024). Selective CDK7 Inhibition Suppresses Cell Cycle Progression and MYC Signaling While Enhancing Apoptosis in Therapy-resistant Estrogen Receptor–positive Breast Cancer. Clinical Cancer Research, 30(9), 1889-1905.

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