The ferroptosis inducer erastin (S7242; Selleck, Shanghai, China), the apoptosis inhibitor Z-DEVD-FMK (T6005; TargetMol, Wellesley Hills, MA, USA) and the necroptosis inhibitor necrostatin-1 (T1847; TargetMol) were each dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich). DFO (D9533; Sigma-Aldrich) was dissolved in deionized water. Cells were seeded in 96-well plates for viability assay, 24-well plates for reactive oxygen species (ROS) detection and immunofluorescence staining, and 6-well plates for western blot assay. To assess the optimum concentration of erastin, neurons were exposed to the drug at 1, 10, 20 or 50 μM 5 days after seeding, and maintained in the medium for 48 hours. The treatment groups were pretreated with Z-DEVD-FMK, necrostatin-1 or DFO for 12 hours. Then, cells were treated with 50 μM erastin for an additional 48 hours. Phosphate-buffered saline (PBS) and DMSO, 10 μL, were used in the PBS and DMSO groups, respectively, as controls.
Necrostatin 1
Manufactured by Targetmol
Sourced in United States, China
Necrostatin-1 is a small-molecule inhibitor that specifically targets and inhibits the enzymatic activity of the receptor-interacting protein kinase 1 (RIPK1). RIPK1 is a key mediator of necroptosis, a form of programmed cell death. Necrostatin-1 has been used in research to study the role of necroptosis in various cellular and biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Chloroquine, by Targetmol (2 mentions)
Z vad fmk, by Targetmol (2 mentions)
Ferrostatin 1, by Targetmol (2 mentions)
Z devd fmk, by Targetmol (1 mentions)
Erastin, by Selleck Chemicals (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Ara c, by Solarbio (1 mentions)
Dimethyl sulfoxide (dmso), by AppliChem (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Gpx4 67763 1 ig, by Proteintech (1 mentions)
Ab72139, by Abcam (1 mentions)
Ab209384, by Abcam (1 mentions)
Ab184718, by Abcam (1 mentions)
Ab187091, by Abcam (1 mentions)
Cycloheximide (chx), by Targetmol (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
4 protocols using necrostatin 1
1
Ferroptosis Induction and Inhibition Assay
2
Investigating Aprepitant's Effects on HL60 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human acute myeloid leukemia cell line HL60 cells were obtained from the Chinese Academy of Medical Sciences. Ara-C (Cat# C8040) was purchased from Solarbio and dissolved in PBS. Aprepitant (Cat# YZ-1041904) was purchased from Solarbio and dissolved in DMSO (Applichem, Cat# A3672) as the stock solution, while the content of DMSO in working solution is less than 0.1%. Z-VAD-FMK (Cat# HY-16658) was purchased from MCE. Necrostatin-1 (Cat# T1847) was purchased from TargetMol. Cells were cultured in RPMI medium 1640 (Gibco, Cat# 31800–022).
3
Multiparametric Analysis of Cell Death
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibodies to PARP (66520-1-Ig), Ki67 (27309-1-AP), NPM1 (10306-1-AP), and GPX4 (67763-1-Ig) were purchased from Proteintech. The antibodies to GPX4 (A1933), GAPDH (AC001), p-NPM1 Thr199 (AP0836), Bax (A19684), Bcl2 (A0208), and YAP1 (A1002) were purchased from ABclonal. The antibody to AURKA (#14475) was purchased from Cell Signaling Technology. The antibodies to p-NPM1 Ser125 (AF3740), p-NPM1 Ser4 (AF8496), and p-NPM1 Thr95 (AF2372) were purchased from Affinity Biosciences. The small-molecule library, TCS7010, ZVAD-FMK, necrostatin-1, ferrostatin-1, and chloroquine were purchased from TargetMol.
4
Investigating Apoptosis and Necroptosis Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies against GAPDH (#97166S, 1 : 1000), PARP (#9542S, 1 : 1000), caspase-3 (#9662S, 1 : 1000), cleaved caspase-3 (#9664S, 1 : 1000), caspase-8 (#9746S, 1 : 1000), cleaved caspase-8 (#9496S, 1 : 1000), Bax (#5023S, 1 : 1000), Bcl-2 (#15071S, 1 : 1000), p-RIP1 (Ser166; #65746S, 1 : 1000), RIP3 (#95702S, 1 : 1000), anti-rabbit IgG, HRP-linked antibody (#7074S, 1 : 3000), and anti-mouse IgG, HRP-linked antibody (#7076S, 1 : 3000) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against MLKL (#ab184718, 1 : 1000), p-MLKL (Ser358; #ab187091, 1 : 1000), RIP1 (#ab72139, 1 : 1000), and p-RIP3 (Ser227; #ab209384, 1 : 1000) were purchased from Abcam (Cambridge, UK). Cladribine, homoharringtonine, Z-VAD-FMK, cycloheximide, chloroquine, ferrostatin-1, and necrostatin-1 were purchased from TargetMol (Shanghai, China). Aclarubicin was obtained from APExBIO (Houston, TX). A stock solution containing cladribine, homoharringtonine, and aclarubicin was prepared in DMSO and kept at -80°C. TNF-alpha was purchased from ABclonal (Wuhan, China).
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!