Hidef detection hrp polymer system

Manufactured by Cell Marque

Sourced in United States

The HiDef Detection HRP Polymer System is a laboratory equipment product designed for immunohistochemistry applications. It serves as a detection system, utilizing a polymer-based approach to amplify and visualize target antigens in tissue samples.

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Lab products found in correlation

Anti mmp2 10 373 2 ap, by Proteintech (1 mentions) Ab244332, by Abcam (1 mentions) Ab955, by Abcam (1 mentions) Liquid dab substrate, by Agilent Technologies (1 mentions) Organosilane adhesive, by Merck Group (1 mentions) Anti upa sc 14019, by Santa Cruz Biotechnology (1 mentions) Trilogy solution, by Cell Marque (1 mentions) Ab92381, by Abcam (1 mentions) Ab6671, by Abcam (1 mentions) Ab218426, by Abcam (1 mentions) Smad3, by Thermo Fisher Scientific (1 mentions) Cleaved cas 3, by Cell Signaling Technology (1 mentions) Katushka, by Evrogen (1 mentions) F4 80, by Abcam (1 mentions) Tunel kit, by MBL Life Science (1 mentions) Dab kit, by Cell Marque (1 mentions)

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HiDef HRP Polymer System (3 citations)

9 protocols using hidef detection hrp polymer system

Immunohistochemistry and Protein Analysis

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Immunohistochemistry staining on paraffin sections was performed with the following primary antibodies: anti-FAPa (NBP2-66844, 1:100; Novus Biologicals, Centennial, CO), anti-CXCL12 (MAB350-100, 1:50; R&D Systems, Herzliya, Israel), and antiepro-CollagenXI-a1 (C10011, 1:100; Oncomatryx, Bizkaia, Spain). HiDef Detection HRP Polymer System (954D-10, Cell Marque, Rocklin, CA) was used as the secondary antibody.
Protein immunoblotting was performed with anti-MMP2 (10373-2-AP, Proteintech, Rosemont, IL) and anti-b-actin (8691001, MP Biomedicals, Auckland, NZ) antibodies.
MF-Fs and NF-Fs medium supernatants were analyzed for CXCL12 by Human SDF-1a ELISA kit (DSA00, R&D Systems), according to the manufacturer's instructions. In brief, samples were diluted 10-fold and 20-fold, incubated for 2 hours, washed, and read in a microplate ELISA reader at 450 nm wavelength.

Aronovich A., Moyal L., Gorovitz B., Amitay-Laish I., Naveh H.P., Forer Y., Maron L., Knaneh J., Ad-El D., Yaacobi D., Barel E., Erez N, & Hodak E. (2021). Cancer-Associated Fibroblasts in Mycosis Fungoides Promote Tumor Cell Migration and Drug Resistance through CXCL12/CXCR4. The Journal of investigative dermatology, 141(3).

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Immunohistochemistry of Pituitary Tumors

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Paraffin-embedded, formalin-fixed tissue blocks were stained with hematoxylin–eosin and reviewed by a pathologist. Tumors were represented with a 2-fold redundancy. Sections (3 μm) were cut and placed onto coated slides. Immunostaining was performed by means of the HiDef detection HRP polymer system (Cell Marque, CA, USA), using specific antibodies against each pituitary hormone (TSH, GH, PRL, FSH, LH and ACTH) and the lineage-specific transcription factors TBX19, POU1F1 and NR5A1, as previously described [54 (link)]. Two independent observers performed assessment of hormones and transcription factors expression at different times.

Andonegui-Elguera S., Silva-Román G., Peña-Martínez E., Taniguchi-Ponciano K., Vela-Patiño S., Remba-Shapiro I., Gómez-Apo E., Espinosa-de-los-Monteros A.L., Portocarrero-Ortiz L.A., Guinto G., Moreno-Jimenez S., Chavez-Macias L., Saucedo R., Basurto-Acevedo L., Lopez-Felix B., Gonzalez-Torres C., Gaytan-Cervantes J., Ayala-Sumuano J.T., Burak-Leipuner A., Marrero-Rodríguez D, & Mercado M. (2022). The Genomic Landscape of Corticotroph Tumors: From Silent Adenomas to ACTH-Secreting Carcinomas. International Journal of Molecular Sciences, 23(9), 4861.

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Immunohistochemical Analysis of CC Lesions

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To investigate the cellular composition and immune status of the CC lesions, 69 samples (36 R and 33 NR patients) were submitted to IHC. Formalin-fixed paraffin-embedded (FFPE) blocks were cut at 4 µm thickness and sections were stained with H&E and IHC. For manual IHC, sections were incubated overnight at 4 °C with the following anti-human antibodies: anti-CD8, rabbit monoclonal (1:500, SP16, Cell Marque, Rocklin, USA); anti-CD4, rabbit monoclonal (1:200, SP35, Cell Marque, Rocklin, USA); anti-PD-1, mouse monoclonal (1:300, ab52587, clone NAT105, Abcam, Cambridge, MA); anti-PD-L1, rabbit monoclonal (1:500, ab205921, clone 28-8, Abcam, Cambridge, MA); anti-PD-L2, rabbit polyclonal (1:500, ab244332, Abcam, Cambridge, MA), anti-CD68, mouse monoclonal (1:100, ab955, Abcam, Cambridge, MA) and anti-FOXP3, rabbit polyclonal antibody (1:1000, PA1-16876, Thermo Fisher Scientific, Rockford, USA). The slides were stained with the Hidef Detection HRP Polymer System (Cell Marque, Rocklin, CA, USA) and visualized with 0.03% 3-3′-diaminobenzidine (SIGMA) in 0.01 M PBS, pH 7.4. A negative control without primary antibodies was generated for each sample.

Rocha Martins P., Luciano Pereira Morais K., de Lima Galdino N.A., Jacauna A., Paula S.O., Magalhães W.C., Zuccherato L.W., Campos L.S., Salles P.G, & Gollob K.J. (2023). Linking tumor immune infiltrate and systemic immune mediators to treatment response and prognosis in advanced cervical cancer. Scientific Reports, 13, 22634.

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Immunohistochemical Detection of Fibrinolytic Factors

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Histological sections (3 μm) were obtained from the paraffin-embedded specimens and mounted on glass slides with organosilane adhesive (3-aminopropyltriethoxysilane; Sigma Chemical Co., St. Louis, USA). For deparaffinization, rehydration, and antigen retrieval, the sections were immersed in Trilogy solution (1:100; Cell Marque, Rocklin, USA) in a Pascal pressure cooker. After these steps, endogenous peroxidase was blocked by treatment with 3% hydrogen peroxide. The sections were incubated overnight in a moist chamber with the following primary antibodies: anti-uPA (sc-14019, 1:800, Santa Cruz Biotechnology, Dallas, USA), anti-uPAR (E-3, 1:200, Santa Cruz Biotechnology, Dallas, USA), and anti-PAI-1 (C-9, 1:400, Santa Cruz Biotechnology, Dallas, USA). The sections were then rinsed twice in phosphate-buffered saline (PBS) and incubated with the HiDef Detection HRP Polymer System (Cell Marque, Rocklin, USA) at room temperature. The reactions were developed using diaminobenzidine (Liquid DAB + Substrate; Dako, Carpinteria, USA) as chromogen. Finally, the sections were counterstained with Mayer's hematoxylin and coverslipped. Endothelial cells present in the histological sections were used as positive internal control. Sections in which the primary antibodies were omitted served as negative control.

Costa C.S.O., Mafra R.P., Rolim L.S.A., Souza L.B, & Pinto L.P. (2022). Immunohistochemical study of the plasminogen activator system in benign epithelial odontogenic lesions. Brazilian oral research, 36.

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Immunohistochemical Evaluation of Cytokines

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Tissues were fixed for one day in a mixture of 2% formaldehyde and 0.2% picric acid in 0.1 M phosphate buffer (pH 7.2). Afterwards, tissues were rinsed in thyroid buffer containing 10% saccharose for 12 h. Samples were embedded into paraffin. Three micrometer thick sections were cut and stained with hematoxylin and eosin (H&E) for subsequent evaluation.
The HiDef Detection HRP Polymer system (Cell MARQUE) was used for the detection of IFN-γ (code ab218426, rabbit polyclonal antibody, working dilution 1:500, Abcam, Cambridge, UK), TNF-α (code ab6671, rabbit polyclonal antibody, working dilution 1:200, Abcam, Cambridge, UK), IL-2 (code ab92381, rabbit monoclonal antibody, working dilution: 1:250, Abcam, Cambridge, UK), IL-7 (code orb13506, rabbit polyclonal antibody, working dilution 1:100, Biorbyt, St. Louis, MO, US), IL-12 (code ab10894, rabbit polyclonal antibody, working dilution 1:200, Biorbyt, St. Louis, MO, US). IL-13 (code ob10895, rabbit polyclonal antibody, working dilution 1:200, Biorbyt, St. Louis, MO, US), and anti-macrophage inflammatory protein 1 beta (MIP-1ß) (ab9675, working dilution 1:100, Abcam, Cambridge, UK).

Reiser S.C., Tellermann J., Akota I, & Pilmane M. (2021). Profiling and Characterization of Localized Cytokine Response in Congenital Cleft Affected Lip Tissue. Life, 11(6), 556.

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Immunohistochemical Assessment of Vascular Type V Collagen and CD34

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The expression of vascular type V collagen and CD34 was assessed immunohistochemically. For this purpose, 4–5 µm-thick FFPE sections mounted on SuperFrost Plus slides (Gerhard Menzel GmbH, Braunschweig, Germany) were used. The IHC protocol recommended by the manufacturer was applied. Briefly, deparaffinized sections were incubated overnight with the primary mouse monoclonal anti-collagen type V antibody (Abcam, Cambridge, UK, 1:300 dilution, ab201980) and mouse monoclonal anti-CD34 antibody (Invitrogen, Camarillo, CA, USA, 1:200 dilution, BI-3C5) at 4 °C. A HiDef Detection HRP Polymer system and a diaminobenzidine tetrahydrochloride substrate kit (Cell Marque, Rocklin, CA, USA) were used to visualize the products of IHC reactions. Cell nuclei were counterstained with Mayer’s hematoxylin. Primary antibodies were omitted in the negative controls of IHC reactions. The reaction results were assessed by two independent observers blind to the clinical data.

Fišere I., Groma V., Svirskis Š., Strautmane E, & Gardovskis A. (2023). Evaluation of Clinical Manifestations of Hemorrhoidal Disease, Carried Out Surgeries and Prolapsed Anorectal Tissues: Associations with ABO Blood Groups of Patients. Journal of Clinical Medicine, 12(15), 5119.

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Immunohistochemical Detection of Leishmania

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The slides were submitted to deparaffinization, rehydration, blocking of endogenous peroxidase, antigen retrieval, blockade of nonspecific protein binding, and incubation with polyclonal rabbit anti‐Leishmania serum diluted 1:500 following a previously described protocol.25 A polymer‐based detection system (HiDef Detection HRP Polymer System; Cell Marque, Rocklin, CA) was used for the detection of amastigote forms of Leishmania spp. according to the manufacturer's recommendations.
For the evaluation of the parasite load in the thymus, macrophages parasitized with amastigote forms of Leishmania spp. were quantified using a 1‐mm² optical grid and a manual cell counter. The cells were counted in five fields at ×400 magnification in the most parasitized areas of the sections. The mean number of parasitized macrophages was calculated and parasitism was classified as absent, mild to moderate (0.2‐10 parasitized macrophages), and intense (more than 10 parasitized macrophages).

da Silva A.V., de Souza T.L., Figueiredo F.B., Mendes AA J.r., Ferreira L.C., Filgueira C.P., Cuervo P., Porrozzi R., Menezes R.C, & Morgado F.N. (2020). Detection of amastigotes and histopathological alterations in the thymus of Leishmania infantum‐infected dogs. Immunity, Inflammation and Disease, 8(2), 127-139.

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Multimodal Analysis of Immune Responses

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Preparation of sections, immunohistochemistry, confocal microscopy, apoptosis TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling) assay and image analysis were performed as previously described [29 (link)]. Lungs and spleens were immune-stained with anti-DTA (Cat#4701, ViroStat, Portland, ME, USA), cleaved Cas-3 (Cat# 9664S, Cell Signaling, MA, USA), P-53 (Cat# SC-6243, Santa Cruz Biotechnology, USA), Mad1 (Cat# 4682, Cell Signaling, MA, USA), E2F1 (Cat# MA5-14344, Thermo Fisher Scientific, Waltham, MA, USA), F4/80(Cat# ab6640, Abcam, Cambridge, MA, USA), Smad3 (Cat# 51-1500, Thermo Fisher Scientific, Waltham, MA, USA), Katushka (Cat# AB233, Evrogen, Moscow, Russia) antibodies at 1:200 dilution and with a TUNEL kit (MBL international, Woburn, MA, USA) according to manufacturer's instructions. Donkey anti-Goat IgG (H+L) secondary antibodies conjugated with Cy2 dye (Cat# 705-225-147, Jackson ImmunoResearch, PA, USA) was used for DTA detection. HiDef Detection HRP Polymer System (Cat# 954D-10, Cell Marque, CA, USA) was used for cleaved Cas-3, P53, Mad1, Smad3, E2F1 and Katushka detection. Pollnk-2 Plus HRP Detection Kit for Rat Primary Antibody (Cat# D46-6, GBI Labs, WA, USA) was used for F4/80 detection.

Elias A., Gritsenko N., Gorovits R., Spector I., Prag G., Yagil E, & Kolot M. (2018). Anti-cancer binary system activated by bacteriophage HK022 integrase. Oncotarget, 9(44), 27487-27501.

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Immunohistochemical Profiling of Melanoma

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Cells isolated from MACS were fixed with a fixation reagent and blocked with a peroxidase blocker (both from Enzo Life Sciences Inc.). Samples were stained using melanoma cocktail: primary antibodies (to HMB-45, MART-1, and tyrosinase), HiDef Detection HRP Polymer System, and a DAB kit (Cell Marque Corporation). Cells were counterstained with hematoxylin and eosin stain. Brown staining of cells was assessed by light microscopy.
In a separate experiment, the samples were stained with CD45-VioBlue and MCSP-APC to detect the presence of leukocytes and melanoma cells, respectively. Samples were visualized using transmission and fluorescent microscopy with various exposure times.

Galanzha E.I., Menyaev Y.A., Yadem A.C., Sarimollaoglu M., Juratli M.A., Nedosekin D.A., Foster S.R., Jamshidi-Parsian A., Siegel E.R., Makhoul I., Hutchins L.F., Suen J.Y, & Zharov V.P. (2019). In vivo liquid biopsy using Cytophone platform for photoacoustic detection of circulating tumor cells in patients with melanoma. Science translational medicine, 11(496), eaat5857.

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