ApopTag Peroxidase In Situ Apoptosis Detection Kit S7100 (Millipore, USA) was used to indicate apoptotic cells in accordance with manufacturer's instructions on 5 μM serial sections. Imaging was performed using Zeiss Axio Scan Z1. Systematic random sampling [40] (link) was utilised to count positive cells against total cell count in the endocardial cushions, primordial MV and TV septal and lateral leaflets to calculate proportions of apoptotic cells for statistical analysis (see below). A total of 224,749 cells were identified as either apoptosis ‘positive’ or ‘negative’ in OFT-banded hearts at HH29 (n = 5 per group).
Apoptag peroxidase in situ apoptosis detection kit s7100
Manufactured by Merck Group
Sourced in United States
The ApopTag Peroxidase In Situ Apoptosis Detection Kit S7100 is a laboratory tool used for the detection and visualization of apoptotic cells in tissue sections and cell populations. It utilizes an enzymatic labeling method to identify DNA fragmentation, a hallmark of programmed cell death. The kit provides the necessary reagents and instructions to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
Axio scan z1, by Zeiss (2 mentions)
Oil red o color solution, by Merck Group (1 mentions)
Cobas 8000 modular analyzer, by Roche (1 mentions)
Axio imager a1, by Zeiss (1 mentions)
Axiocam mrc camera, by Zeiss (1 mentions)
Alexa fluor 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Hoechst 33432, by Thermo Fisher Scientific (1 mentions)
Anti f4 80, by Bio-Rad (1 mentions)
Pcna sc 56, by Santa Cruz Biotechnology (1 mentions)
Dmc2900 camera, by Leica camera (1 mentions)
Pi 2000 1, by Vector Laboratories (1 mentions)
Entellan new, by Merck Group (1 mentions)
Safranin o, by Merck Group (1 mentions)
Fluorescent microscope, by Nikon (1 mentions)
Image pro plus 6, by Nikon (1 mentions)
Np 40, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Proteinase k, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Na2hpo4, by Merck Group (1 mentions)
22 protocols using apoptag peroxidase in situ apoptosis detection kit s7100
1
Apoptosis Detection in Cardiac Development
2
Immunohistochemical Analysis of Proliferation, Apoptosis, and Lymphocyte Infiltration
3
TUNEL Assay for Apoptosis Detection
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The fragmentation of DNA was examined using an ApopTag® Peroxidase in situ Apoptosis Detection Kit (S7100) (Millipore) according to the manufacturer's instructions. Briefly, brain sections from each mouse (n = 5 per group) were subjected to enzymatic digestion with 20 μg/ml proteinase K for 5 min, treated with 5% hydrogen peroxide for 20 min to exhaust endogenous peroxidase activity, and washed with PBS. They were then immersed in ApopTag® equilibration buffer for 10 min to label the 3′-OH ends of fragmented DNA and incubated with terminal deoxynucleotidyl transferase-mediated UTP nick end labeling (TUNEL) at 37°C for 1 h. After being washed with PBS, the sections were incubated with DAB to detect signs of apoptotic cell death.
4
Histological Analysis of Apoptosis
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After the PET scans, hearts were excised. After fixation in 4% phosphate-buffered formalin, hearts were cut into 4-µm-thick sections and embedded in paraffin. Standard histological procedure (Haematoxyline-Eosine and Giemsa staining) was performed. According to the manufacturer's protocol, apoptotic cells were stained using ApopTag® Peroxidase In Situ Apoptosis Detection Kit S7100 (Millipore) and evaluated by the Axio Version SE64 (Version 4.9) software.
5
Assessing Testicular Apoptosis
6
Histological Analysis of Liver Lipids and Apoptosis
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Livers were fixed overnight in 4% paraformaldehyde, embedded in paraffin, and sectioned at a thickness of 6 μm. For Oil Red O staining, 10‐µm liver cryosections were stained with commercially available kits (Oil Red O color solution, Cat# 1.20419.0250; Merck, Germany). Terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick‐end labeling (TUNEL) staining was performed using the ApopTag Peroxidase In Situ Apoptosis Detection Kit (S7100; Millipore, Temecula, CA).
7
Toxicity Evaluation of KOK in Prepuberal Rats
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To investigate whether KOK has a toxic effect for long-term use in rats, KOK (2.0 and 4.0 g/kg/day) was orally administrated to normal female prepuberal rats (23-days-old) for 22 days. Body weight was measured and serum was obtained at 24 hours after the last administration of KOK. Serum level of alanine aminotranferease (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) was measured using an enzymatic or ultraviolet assay with ALT, AST, or LDH detection kit (Roche, Basel, Switzerland) according to the manufacturer's instructions under cobas 8000 modular analyzer (Roche, Basel, Swetzerland). General structure, apoptotic cell death, and proinflammatory cytokines were evaluated by H&E staining, TUNEL staining, immunohistochemistry, and Western blot analysis for capase-3, or real time PCR analysis for TNF-α and fatty acid synthase (FAS), according to pre-described protocols and manufacturer's instructions (ApopTag peroxidase in situ Apoptosis Detection Kit, S7100, Millipore, USA).
8
Histopathological Analysis of Tissue Ablation
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Endoscopy confirmed the absence of bleeding or perforation in the lesions at 24 h post-ablation and then pigs were sacrificed. After the electroporated tissues were fixed in 10% formalin solution for 24 h. Next, the fixed tissues were mounted in paraffin and sliced into 3-µm-thick sections and analyzed using hematoxylin and eosin (H&E) staining. An additional TUNEL assay (ApopTag® Peroxidase In Situ Apoptosis Detection Kit, S7100, Millipore, Gaithersberg, MD, USA) was performed to assess tissue apoptosis. After fixation, the presence of brown stained cells was considered positive TUNEL assay findings.
9
TUNEL Staining for Apoptosis Detection
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To evaluate apoptosis, TUNEL staining was performed using the Apoptag® Peroxidase In Situ Apoptosis Detection Kit S7100 (Millipore Corp., Billerica, MA, USA).19 (link) After deparaffinization with xylene and graded concentrations of alcohol, gastric mucosal tissue sections were exposed to Proteinase K for 15 minutes at room temperature. Endogenous peroxidase activity was quenched with Inhibitor D for 5 minutes at room temperature. Sections were incubated with terminal deoxynucleotidyl transferase (TdT) in a humidified chamber at 37°C for 1 hour. After incubation with anti-digoxigenin-conjugate for 30 minutes at room temperature, peroxidase substrate and 0.05% DAB was applied to develop color. The specimens were then washed with distilled water and counterstained with 0.5% methyl green for 10 minutes at room temperature. Sections were counterstained with Mayer’s hematoxylin. Identically-treated slides not exposed to TdT served as negative controls. In all cases, positive staining was qualitatively evaluated on the entire slide by an experienced pathologist (E.S.).
10
Apoptosis Detection Using TUNEL Assay
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The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed using an ApopTag Peroxidase in situ Apoptosis Detection Kit (S7100) (Millipore, Burlington, MA) according to the manufacturer's instructions and as previously described [23] (link).
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