An in situ cell death detection peroxidase (POD) kit (Roche Diagnostics, Mannheim, Germany) was used for the TUNEL technique and the sections were stained according to the manufacturer’s protocol. In each sampling frame, a counting area was designated without bias and the apoptotic index was calculated by dividing the apoptotic (TUNEL-positive) RGC number by the total RGC number [19]. TUNEL-positive RGCs were counted by the same investigator (UU) blinded to the group assignment.
Pod kit
Manufactured by Roche
Sourced in Germany
The POD Kit is a laboratory equipment product from Roche. It serves as a core component for various applications. The POD Kit provides a functional platform to support specific laboratory processes.
Automatically generated - may contain errors
7 protocols using pod kit
1
TUNEL Assay for Apoptotic RGCs
2
TUNEL Assay for Apoptosis Quantification
3
Liver Histology and Apoptosis Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The liver was cut into 4 μm frozen sections and stored at −20°C. Next, the sections were fixed in 4% paraformaldehyde for 30 min at room temperature, stained with 0.3% oil red O solution for 30 min, counter-stained with hematoxylin and eosin (H&E) for 30 s, and sealed with glycerin. Liver lipid accumulation was evaluated using an optical microscope (200×).
In addition, liver tissues were fixed in 4% paraformaldehyde for 12–24 h, dehydrated in absolute ethanol, transparentized in dimethylbenzene, embedded in paraffin, sectioned into 4 μm sections, deparaffinized, dehydrated with gradient ethanol solutions, routinely stained with H&E, and sealed with optical resin. Pathological changes of the liver tissues were observed using an optical microscope (200×).
Using POD kit (Roche, Basel, Switzerland), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was conducted according to the manufacturer’s guidelines. Hepatocytes that showed a positive TUNEL staining were observed using an optical microscope (200×). In each section, 5 random fields were selected and the number of apoptotic cells per field was counted using Image-Pro Plus 5.1 software (Media Cybernetics, Rockville, MD, USA).
In addition, liver tissues were fixed in 4% paraformaldehyde for 12–24 h, dehydrated in absolute ethanol, transparentized in dimethylbenzene, embedded in paraffin, sectioned into 4 μm sections, deparaffinized, dehydrated with gradient ethanol solutions, routinely stained with H&E, and sealed with optical resin. Pathological changes of the liver tissues were observed using an optical microscope (200×).
Using POD kit (Roche, Basel, Switzerland), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was conducted according to the manufacturer’s guidelines. Hepatocytes that showed a positive TUNEL staining were observed using an optical microscope (200×). In each section, 5 random fields were selected and the number of apoptotic cells per field was counted using Image-Pro Plus 5.1 software (Media Cybernetics, Rockville, MD, USA).
4
TUNEL Assay for Cell Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To investigate the cell apoptosis, an in situ cell death detection POD Kit (Roche) was used for the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) technique and the sections were stained according to the manufacturer’s instructions for paraffin-embedded tissues. We visualized the POD retained in the immune complex with a DAB detection kit (DAKO). Sections were assessed using an optical microscope (Olympus).
5
Quantitative Analysis of Retinal Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was detected with the cell death detection POD Kit (Lot#11684795910, Roche Diagnostics GmbH, Mannheim, Germany). Briefly, the rat retinal sections were prepared for TUNEL assay according to the instruction. The sections were counterstained with DAPI (#C1005, Beyotime, Shanghai, China) and then analyzed under the digital immunofluorescence microscope imaging system (DP71, Olympus, Japan). The number of TUNEL-positive cells among the ONL was counted in 3 sections from each rat retina and was averaged.
6
Quantifying Retinal Cell Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Terminal deoxyuridine triphosphate nick-end labeling (TUNEL) assay was performed to detect the apoptotic cells in retinal sections following a previous described method (Yoshizawa et al., 2000 (link)). An in situ cell death detection POD Kit (Roche, Mannheim Germany) was used according to manufacturer’s protocol. TUNEL sections were counterstained with DAPI, mounted on slides, and then visualized with confocal microscopy (LSM510, Zeiss, Oberkochen, Germany). Apoptotic index (AI) of the ONL was calculated on the basis of cell numbers (number of TUNEL-positive nuclei/total number of photoreceptor cell nucleix100).
7
Quantifying Intestinal Epithelial Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues were fixed in 4% paraformaldehyde, and 10-μm frozen sections were prepared. Intestinal epithelial apoptosis was investigated using terminal deoxynucleotidyl transferase mediated dUTP nickend labeling (TUNEL) staining and an in situ cell death detection POD kit (Roche, Penzberg, Germany) in accordance with the manufacturer's instructions. The slides were rinsed with phosphate-buffered saline (PBS) for 10 min at room temperature. Slides were incubated with 0.1% Triton X-100 for 8 min, rinsed with PBS for 15 min and 50 μL of the TUNEL reaction mixture was added and incubated for 60 min at 37 °C. Slides were rinsed three times with PBS and examined under a fluorescence microscope. The ratio of TUNEL-positive cells was calculated as follows: (number of apoptotic cells/total number counted) ⨉100%. Each assay was performed in a blinded manner, and the experiment was repeated three times.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!