The OC tissue specimens resected from 45 patients (date range: 04/2015~03/2019; age distribution: 39~75) at Kobe University Hospital (Kobe, Japan) were fixed with 10% (v/v) formalin at room temperature for 48 h and embedded in paraffin. Cylindrical tissue cores (2 mm) were then extracted from each paraffin block and re-embedded into a single paraffin block to create a tissue microarray (TMA) for sectioning. Epithelial OC cases were selected from the TMA and classified into platinum-sensitive and platinum-resistant groups according to their clinical course. The resultant TMA sections were incubated with antibody against Fdx1 (1:200; cat. no. 12592-1-AP; Thermo Fisher Scientific, Inc.) overnight at 4°C and then with anti-Rabbit IgG antibodies conjugated with HRP-labeled polymer (ImmPRESS Reagent kit Peroxidase; Vector Laboratories, Inc.) for 30 min at room temperature. Secondary antibodies were visualized using DAB Chromogen (Dako; Agilent Technologies, Inc.), and nuclei were counterstained with hematoxylin. The specimens were observed under a BZ-X700 microscope (Keyence Corporation). Clinical tissue specimens were obtained by opt-out method and analyzed in accordance with procedures approved (approval nos. B200076 and B220122) by the Institutional Review Board of Kobe University Hospital (Kobe, Japan).
Immpress reagent kit peroxidase
Manufactured by Vector Laboratories
The ImmPRESS Reagent kit Peroxidase is a laboratory product designed for use in immunohistochemistry applications. It contains a secondary antibody conjugated to a peroxidase enzyme, which can be used to detect primary antibodies bound to target antigens in tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Dab chromogen, by Agilent Technologies (1 mentions)
Bz x700 microscope, by Keyence (1 mentions)
Ab24261, by Abcam (1 mentions)
Dab reagent, by Vector Laboratories (1 mentions)
Dm1000 light microscope, by Leica (1 mentions)
Application suite software version 3, by Leica (1 mentions)
Sc 49480, by Santa Cruz Biotechnology (1 mentions)
Haematoxylin, by Merck Group (1 mentions)
Ab5694, by Abcam (1 mentions)
2 protocols using immpress reagent kit peroxidase
1
Fdx1 Expression in Ovarian Cancer
2
Immunohistochemical Analysis of Tissue Markers
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Immunohistochemistry was performed as previously described15 (link),21 (link),34 (link). In brief, tissues were deparaffinized and immune-labeled using primary antibodies against periostin (sc49480; Santa Cruz Biotechnology; Dallas, TX; 1:100), α-SMA (ab5694; Abcam; Cambridge, MA; 1:400), PCNA (ab2426-1; Abcam; 1:100), and FSP-1 (07-2274; Millipore; Billerica, MA; 1:100). Primary antibodies were detected using the ImmPRESS Reagent Kit Peroxidase (Vector Laboratory; Burlingame, CA) and DAB reagent (Vector Laboratory) following the manufacturer’s instructions. Primary antibody delete was used as experimental control. All sections were counterstained with haematoxylin (Sigma Aldrich). To study collagen levels, deparaffinized histological sections were stained using Masson’s trichrome (University Hospital, London, ON) stain. Images were taken with a DM1000 light microscope (Leica; Concord, Ontario) and Leica Application Suite Software (version 3.8).
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