To further evaluate the effect of (±)-RPE65-61 on retinal apoptosis, retinal sections from LIRD mice were used for TUNEL with the In Situ Cell Death Detection Kit (Millipore/Sigma Cat#:11684795910). TUNEL-positive photoreceptor cells were quantified to evaluate the effects of (±)-RPE65-61 as described [29 (link)].
In situ cell death detection kit
Manufactured by Merck Group
Sourced in United States, Germany
The In Situ Cell Death Detection Kit is a laboratory product designed to detect and analyze cell death in various biological samples. It provides a sensitive and reliable tool for researchers to assess apoptosis, necrosis, and other forms of programmed cell death during different experimental conditions.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (9 mentions)
Trypsin edta, by Thermo Fisher Scientific (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
Alexa fluor 594 conjugated wheat germ agglutinin, by Thermo Fisher Scientific (3 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (3 mentions)
Caspase 3, by Cell Signaling Technology (3 mentions)
Bcl 2, by Cell Signaling Technology (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Trizol, by Thermo Fisher Scientific (3 mentions)
Lsm 700, by Zeiss (3 mentions)
Hoechst 33342, by Thermo Fisher Scientific (3 mentions)
Propidium iodide, by Thermo Fisher Scientific (3 mentions)
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Axio observer z1, by Zeiss (3 mentions)
Fluorescein, by Merck Group (2 mentions)
Tmr red, by Merck Group (2 mentions)
Oil red o, by Merck Group (2 mentions)
Oxaliplatin, by Merck Group (2 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
164 protocols using in situ cell death detection kit
1
Evaluating RPE65-61 Effects on Retinal Apoptosis
2
Histological Analysis of Testes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The testes were fixed with Davidson’s fixative for 24 h at room temperature and embedded in paraffin. Samples were cut and stained with HE. TUNEL staining was performed using the In Situ Cell Death Detection Kit (MilliporeSigma) according to the manufacturer’s instructions. Images were obtained using the Leica TCS SP8 Laser Scanning Confocal Microscope (Leica, Wetzlar, Germany).
3
Endothelial Cells Morphology and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HCAECs at a density of 18,000 cells/well or MPLECs at 25,000 cells/well were seeded onto 24-well plates. HCAECs were treated with or without EndMT and with or without MC1568 as described above. MPLECs were treated with 4-OH tamoxifen or vehicle and with or without EndMT, as described above (quantities of reagents and media were scaled to 24-well format). Cells were then fixed with ice-cold 4% PFA for 10 minutes and stained with 0.5% crystal violet (C3886, MilliporeSigma) for another 10 minutes before washing with tap water. Images were taken with an inverted microscope (DMi8, Leica). For TUNEL assay, HCAECs or MPLECs were cultured and fixed as described for the crystal violet assay. An In Situ Cell Death Detection Kit (12156792910, MilliporeSigma) was used according to the manufacturer’s instructions.
4
In Situ Cell Death Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested, fixed in 4% paraformaldehyde (MilliporeSigma) at room temperature for 15 min and permeabilized with 0.25% Triton-X100 (Dalian Meilun Biotechnology Co., Ltd.). Thereafter, cells were stained using an in situ cell death detection kit (MilliporeSigma). Images were visualized using a fluorescence microscope (Nikon Corporation). The cell death rate was calculated as TUNEL-positive cells/total cells ×100.
5
TUNEL Assay for Cell Death
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryosections were analyzed with an In Situ Cell Death Detection Kit (Millipore-Sigma). Slides were washed three times in PBS for 10 min and permeabilized in PBS with 0.1% Triton-X-100 for 2 min on ice. Sections were washed twice in PBS for 10 min and incubated with 50 µL of TUNEL reaction mixture for 60 min at 37°C. Slides were washed three times for 10 min with PBS and incubated with 1.6 μmol/L Hoechst nuclear stain and imaged by confocal laser microscopy (λex = 488 nm) with frame-stack sequential scanning.
6
TUNEL Assay for Embryo Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryos were fixed, permeabilized, and blocked as described for immunofluorescence. Zonae pellucida were removed using Tyrode’s Acid treatment prior to performing the TUNEL assay (In Situ Cell Death Detection Kit, Fluorescein, Millipore-Sigma). Embryos were incubated in 200 µl of a 1:10 dilution of enzyme in label solution for 2 hr at 37°C. Embryos were then washed twice with blocking solution for 10 min each, and then mounted in a 1 to 400 dilution of DRAQ5 in blocking solution to stain DNA.
7
Intestinal Tissue Cryoprotection and Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After flushing with cold PBS, small intestinal tissues were incubated overnight in 30% sucrose in PBS for cryoprotection. Six mm-thick cryostat sections were prepared and stored at −80 °C until needed. The frozen sections were fixed with ice-cold 100% ethanol and acetone at the ratio of 1:1 for 10 min at −20 °C. EdU-labeled cells were stained using the Click-iT EdU Cell Proliferation kit (Thermo Fisher). Terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining was performed on intestinal tissues and enteroids using an in situ cell death detection kit (MilliporeSigma, St. Louis, MO, USA) according to the manufacturer’s instruction. Images were taken using a Nikon A1R HD confocal microscope (Nikon Instruments, Melville, NY, USA).
8
TUNEL Assay for Apoptosis Detection
9
Quantifying Apoptosis in Colon Epithelium
10
Apelin and Sirt3 Modulate Mitochondrial Homeostasis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Icariin (purity 98.75%) and JC-1 dye were purchased from MedChem Express (MCE, NJ, USA). The dihydroethidium (DHE) probe and mitochondria isolation kits were obtained from Beyotime Biotechnology (Jiangsu, China). The in situ cell death detection kit (Roche) was obtained from Sigma-Aldrich (St. Louis, MO, USA); and the primary antibodies against Sirt3, Sirt1, PGC-1α, Mfn2, Cyt-b, and β-actin, as well as the goat anti-rabbit and goat anti-mouse secondary antibodies were obtained from Abcam Biotechnology (Cambridge, MA, USA). The primary antibody against Apelin was obtained from Signalway Antibody (SAB, USA). Empty adenoviral vectors (Ad-EV) and recombinant adenoviral vectors expressing Apelin (Ad-Apelin), Sirt3 (Ad-Sirt3) or Apelin-specific small hairpin RNA (Ad-sh-Apelin), as well as Sirt3-specific small hairpin RNA (Ad-sh-Sirt3), were purchased from Hanbio Technology Ltd. (Shanghai, China). The titer of the adenoviruses was approximately 1.2×1010 PFU/mL.
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!