Four-micron sections of formalin-fixed paraffin-embedded mammary tumors were deparaffinized, rehydrated, heated in citrate buffer (pH 6), and stained using antibodies against PgR (Dako; A0089; 1:100). Positive signal was developed according to instructions from the EnVision HRP System (Dako) or MACH 4 (Biocare) followed by DAB chromogen.
Mach 4
Manufactured by Biocare Medical
Sourced in United States
The MACH 4 is a high-performance laboratory instrument designed for automated hematology analysis. It provides accurate and reliable measurement of various blood cell parameters, including red blood cells, white blood cells, and platelets. The MACH 4 utilizes advanced technology to deliver efficient and consistent results, making it a valuable tool for clinical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Envision system hrp, by Agilent Technologies (1 mentions)
Bloxall, by Vector Laboratories (1 mentions)
Background punisher, by Biocare Medical (1 mentions)
Warp red, by Biocare Medical (1 mentions)
Tacha s hematoxylin, by Biocare Medical (1 mentions)
Citrisolve, by Decon Labs (1 mentions)
Ecomount, by Biocare Medical (1 mentions)
Nbp1 42140, by Novus Biologicals (1 mentions)
Ab9498, by Abcam (1 mentions)
Sc 365712, by Santa Cruz Biotechnology (1 mentions)
Ab10558, by Abcam (1 mentions)
Background sniper, by Biocare Medical (1 mentions)
Mirax midi slide scanner, by Zeiss (1 mentions)
Nb100 2284, by Abcam (1 mentions)
Ab5694, by Abcam (1 mentions)
Pannoramic viewer, by 3DHISTECH (1 mentions)
4 protocols using mach 4
1
Immunohistochemical Analysis of Progesterone Receptor in Mammary Tumors
2
Immunohistochemical Staining for Protein Kinase
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Tissue sections, 3–4 μm in thickness, were deparaffinized in CitriSolve (Decon Labs., King of Prussia, PA) and rehydrated by processing them through graded alcohol solutions. Antigen unmasking was accomplished by digesting tissue in 10 μg/ml of protein kinase for 15 min at room temperature (Fisher Scientific, Waltham, MA). Endogenous enzymes and non-specific background were blocked with Background Punisher (Biocare Medical, Pacheco, CA), followed by BLOXALL (Vector Laboratories, Burlingame, CA). The primary antibody (NBP1–42140; Novus Biologicals, Centennial, CO) was incubated on the tissue sections at a dilution of 1:100 for 1 h at room temperature. Subsequently, sections were sequentially incubated with an alkaline phosphatase-based detection polymer kit (MACH 4; Biocare Medical), and Warp Red (Biocare Medical). Sections were counterstained with Tacha’s hematoxylin (Biocare Medical) and mounted using a permanent mounting medium (EcoMount; Biocare Medical).
3
Quantifying Angiogenesis in Matrigel Plugs
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Immunohistochemistry was performed to confirm angiogenesis in the Matrigel plug and to quantify capillary density. First, slides were deparaffinized, hydrated, and treated with hydrogen peroxide 0.3% for 15 minutes. The tissues were then blocked (BSA 10%) and incubated at 4°C with anti‐mouse von Willebrand factor (1:200; sc‐365712; Santa Cruz, Dallas, TX) or anti‐mouse CD‐31 for capillary density (1:100; ab9498; Abcam, Cambridge, UK). Immunodetection was performed using the VECTASTAIN Elite ABC Kit (Agilent (Dako), Santa Clara, CA) or MACH 4 (Biocare Medical, Pacheco, CA), and the antigen‐antibody complex was visualized using diaminobenzidine. Slices were examined using a brightfield microscope (Eclipse 50i and 90i, NIS‐Elements software, Nikon, Tokyo, Japan) by an investigator blinded to the samples. Positive microvessel stains were counted in four random fields of three different sections (n = 6 hearts per group) using a 40x objective and reported as the number of capillaries per square millimeter.
4
Histological Evaluation of Cellulose Scaffold
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Serial 5μm thick sections were cut, beginning at 1 mm inside the cellulose scaffold, and stained with hematoxylin-eosin (H&E) and Masson’s trichrome. For immunocytochemistry, heat induced epitope retrieval was performed at 110°C for 12 min with citrate buffer (pH 6.0). Anti-CD31/PECAM1 (1:100; Novus Biologicals, NB100-2284, Oakville, ON, Canada), anti-alpha smooth muscle actin (1:1000, ab5694, abcam, Toronto, ON, Canada) and anti-CD45 (1:3000; ab10558, abcam, Toronto, ON, Canada) primary antibodies were incubated for a hour at room temperature. Blocking reagent (Background Sniper, Biocare, Medical, Concorde, CA, USA) and detection system MACH 4 (Biocare Medical, Concord, CA, USA) were applied according to company specifications. For the evaluation of cell infiltration, extracellular matrix deposition and vascularisation (angiogenesis), micrographs were captured using Zeiss MIRAX MIDI Slide Scanner (Zeiss, Toronto, Canada) equipped with 40x objective and analysed using Pannoramic Viewer (3DHISTECH Ltd., Budapest, Hungary) and ImageJ software. The scoring of inflammation was evaluated by a pathologist. The scoring was subjectively assigned by qualitative analysis of the magnitude of the total foreign response as well, the cell population proportions within the foreign response.
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