The immunohistochemistry (IHC) staining procedure was optimized as described previously (37 (link)) using the non-biotin-polymerized horseradish peroxidase (HRP) system (BioGenex Laboratories, San Ramon, CA). Briefly, intestinal tissue sections from each pig were deparaffinized and rehydrated in graded ethanol to PBS (pH 7.4). Antigen retrieval and unmasking were performed by treatment with 0.05% pronase E (Sigma-Aldrich, St. Louis, MO) for 20 min. The endogenous peroxidase activity was quenched with 3% hydrogen peroxide (Sigma) for 20 min. Then, the sections were incubated in Power Block solution (BioGenex) for 30 min at room temperature. Mouse monoclonal antibody anti-PEDV nucleocapsid protein (N) (SD6-29; a gift from E. Nelson and S. Lawson, South Dakota State University) was applied to each section at 4°C overnight. After two washes in PBS, a commercial Super Sensitive IHC detection system (BioGenex) was used. Finally, these sections were counterstained with Mayer’s hematoxylin (BioGenex) and dehydrated, and coverslips were added. The IHC signal of PEDV antigen was scored as 0 to 3 according to the percentage of villous enterocytes within the section showing a positive signal. Scores were as follows: 0 indicated that there were no positive cells and 1, 2, and 3 indicated that <30%, 30 to 60%, and >60% of villous enterocytes showed a positive signal, respectively.
Super sensitive ihc detection system
Manufactured by BioGenex
Sourced in United States
The Super Sensitive IHC Detection Systems by BioGenex are a set of reagents and kits designed to facilitate sensitive and specific immunohistochemical (IHC) staining. The core function of these systems is to enhance the detection of target antigens in tissue samples, enabling clear visualization and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Pronase e, by Merck Group (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Mayer s hematoxylin, by BioGenex (1 mentions)
Power block solution, by BioGenex (1 mentions)
Mab1952z, by Merck Group (1 mentions)
Avidin biotin blocking kit, by Vector Laboratories (1 mentions)
Ultravision block, by Thermo Fisher Scientific (1 mentions)
Hydrogen peroxide block, by Thermo Fisher Scientific (1 mentions)
Ab9574, by Abcam (1 mentions)
Ab6002, by Abcam (1 mentions)
Ab253183, by Abcam (1 mentions)
Ab126577, by Abcam (1 mentions)
Sc 166965, by Santa Cruz Biotechnology (1 mentions)
Hydrogen peroxide, by Thermo Fisher Scientific (1 mentions)
Trichrome stain kit, by Abcam (1 mentions)
α sma, by Abcam (1 mentions)
Antigen retrieval citra solution ph 6, by BioGenex (1 mentions)
Blocking buffer, by Roche (1 mentions)
Antigen unmasking solution, by Vector Laboratories (1 mentions)
Hematoxylin, by Muto Pure Chemicals (1 mentions)
11 protocols using super sensitive ihc detection system
1
Optimized Immunohistochemistry Procedure for PEDV
2
Ovarian Cancer Immunohistochemical Analysis
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In total, 31 human ovarian cancer surgical specimens were obtained from the archives of the Department of Pathology, National Taiwan University Hospital. This study was approved by the National Taiwan University Hospital Research Ethics Committee. For IHC, 4 µm paraffin‐embedded sections were stained using the Super Sensitive™ IHC Detection System (BioGenex) according to the manufacturer's directions. Anti‐integrin α3 (MAB1952Z, Merck) and anti‐laminin antibody (ab17107, Abcam) were diluted to 1:100 and incubated with tissue sections overnight. Samples with less than 1% positive cells were considered as negative staining. The intensity of staining in positive cases was graded as weak, moderate, or strong. Staining was evaluated by a pathologist, Dr. Kuo, K‐T.
3
Immunohistochemical Staining Protocol
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Sections of tissue sample were deparaffinized and rehydrated. Antigen retrieval was performed by heating slides in 10 mM citrate buffer (pH 6.0) for 40 min. Endogenous peroxidase activity was inactivated by 3% hydrogen peroxide. The slides were then blocked with 0.005 g/mL bovine serum albumin (BSA) for 30 min, and primary antibodies were added. After incubating with primary antibodies, the sections were examined using the Super Sensitive IHC detection system (Biogenex, CA, USA) with 3-amino-9-ethylcarbazole (AEC) chromogen and counterstained with hematoxylin. Slides in the absence of primary antibody were used as negative controls.
4
Immunohistochemical Analysis of Pancreatic Insulin
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After fixation in 10% formalin for 24 hr, the pancreas tissues were sliced and embedded in paraffin. Then, tissue was sectioned at 3 to 4 µm thickness. The section slide was de-waxed, further sequentially treated with 100, 95, 80, and 70% ethanol, and then heated to activate the antigen. After washing with PBST (phosphate-buffered saline mixed with Tween 20, 100: 0.05), the tissues were stained with a Super Sensitive™ IHC detection system (BioGenex, USA). Then, the signals were revealed by DAB Quanto Chromogen with Substrate (1:30). The insulin positive areas (IPAs) were observed under a microscope. IPA diameters larger than 20 µm and smaller than 20 µm were counted in triplicate on the anterior, median and posterior sides of the pancreas, and the IPA value represented the mean value of 9 counts.
5
Immunohistochemical Staining of Paraffin Sections
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For immunohistochemistry 3 μm paraffin sections were deparaffinized in xylol and re-hydrated in a descending alcohol series. Antigen retrieval was achieved by heating for 15 min at 89 °C in citrate-buffer (pH 6.0). Intrinsic biotin and avidin binding sites were blocked with Avidin-Biotin Blocking Kit (Vector Laboratories, Burlingham, CA), endogenous peroxidase-activity with Hydrogen-Peroxide Block (15 min, RT; Thermo Scientific, Fremont, CA) and unspecific background was reduced with Ultra-Vision Block (5 min, RT; Thermo Scientific, Fremont, CA, USA). Slides were incubated with primary antibodies as previously described (clone TR1.02; Ganten et al., 2009 [23 (link)]). Bound antibodies were detected by a Super Sensitive IHC Detection System (BioGenex, San Ramon, USA). For color development, a Fast Red system (Sigma, Deisenhofen, Germany) was used. Washing steps were done with Tris-buffered saline supplemented with Tween (TBST). All slides were counterstained with hemalum and cover slipped.
6
Immunohistochemical Analysis of LASP-1 Expression
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Tissue sections (5 μm) were de-paraffinized in xylene, rehydrated in ethanol, incubated in 0.3% H2O2 in methanol for 20 min to block endogenous peroxidase activity; 3% BSA was used to block non-specific staining. The sections were washed with 1X PBS and incubated with rabbit anti-human LASP-1 (1:50 v/v) overnight (Santa Cruz Biotechnology, Biosource, CA, USA). The biotinylated secondary antibodies were added for 15 min (Super Sensitive IHC Detection Systems, BioGenex, San Ramon, CA, USA). After extensive washing, the sections were incubated with horseradish peroxidase complex (ABC complex) for 15 min. The chromogen DAB was used to localize the peroxidase in tissues. The slides were counterstained with H&E and analyzed with an optical microscope at ×10, ×40 and ×60 magnification.
7
Cartilage Degeneration and Synovitis Assessment
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Ten sections of knee joints spanning 200 μm were Safranin-O stained. Articular cartilage destruction was evaluated, according to the guideline of Osteoarthritis Research Society International [51 (link)]. Synovitis was graded using a 0–3 scoring system, with 0 = normal, 1 = moderate, and 3 = severe degeneration. Immunoreactivity was examined using Utx (ab253183, Abcam), H3K27me3 (ab6002; Abcam), Tfam (SC166965; Santa Cruz), Igf2 (ab9574; Abcam), Ezh2 (ab283273, Abcam), Eed (ab264566, Abcam), Suz12 (ab126577, Abcam), and Super Sensitive™ IHC Detection Systems (BioGenex Laboratories). Immuno-labeled chondrocytes within each field (50 μm2) were counted (nine fields in three sections for each specimen).
8
Histological Assessment of Pancreatic Fibrosis
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Masson’s trichrome staining was performed on pancreas tissue sections using a Trichrome Stain kit from Abcam, according to the manufacturer’s protocol. For immunohistochemistry, paraffin-embedded pancreas tissue slides were deparaffinized with xylene and hydrated through a graded alcohol series. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide (Fisher Scientific) for 10 min. Antigen retrieval was performed using Antigen Retrieval Citra Solution pH 6 (BioGenex) in a heat cooker for 30 min, followed by avidin/biotin block (Life Technologies) with 10% goat serum + 1% BSA in phosphate-buffered saline and incubated for 30 min. Primary antibody for alpha-smooth muscle actin (α-SMA, 1:200, Abcam) was incubated overnight. Immunostaining was performed using Super Sensitive™ IHC Detection Systems (BioGenex). Slides were counterstained with DAB chromagen. Both Masson’s trichrome and α-SMA stained slides were scanned at 20x using an Aperio AT2 slide scanner. Three fields were randomly selected from five pancreas tissue slides in each group, and the percent positively stained area in each field was determined using Image J software.
9
Immunohistochemical Analysis of RacGAP1
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The tumor tissues were collected and placed in 4% PFA solution, fixed for 24 h, dehydrated through a gradient of ethanol, and embedded into paraffin blocks for immunohistochemistry. The paraffin blocks were cut to 4 µm sections and mounted onto microscope slides for analysis. For antigen retrieval, the slides of tumor sections were incubated with antigen unmasking solution (Vector Laboratories, Burlingame, CA, USA) at 80 °C for 1 h. Endogenous peroxidase activity was quenched using 3% H2O2 in 10% methanol. Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight. Super Sensitive IHC Detection Systems (BioGenex, Fremont, CA, USA) were used to amplify the signal. Sections were stained with horseradish peroxidase (HRP) secondary antibodies. After two washes, the slides were counterstained with hematoxylin (Muto Pure Chemicals, Tokyo, Japan). Positive staining was scored using the following formula: (r3/t) × 3 + (r2/t) × 2 + (r1/t) × 1, where t is the total area of tumor tissue for a whole tumor section, r3 is the total area of high-intensity staining (intensity 3), r2 is the total area of medium intensity staining (intensity 2), and r1 is the total area of weak intensity staining (intensity 1).
10
Skin Tissue Analysis by Histology
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