HepG2 (SCSP-510), Huh7 (SCSP-526), and L02 (GNHu 6) were purchased from the Cell Bank, Chinese Academy of Medical Sciences (Shanghai, China). MHCC97H were obtained from the Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China. HepG2-Luc were constructed by Cloud-Clone Corp (Wuhan, China). All cell lines were authenticated, and cells were thawed upon arrival, expanded, and stored in a liquid nitrogen tank. The absence of mycoplasma cell contamination was confirmed using MycAwayTM-Color One-Step Mycoplasma Detection Kit (40611ES, Yeasen, China). All the cell lines were cultured in Dulbecco’s modified Eagle’s medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA) at 37°C, 5% CO2. Ganetespib (STA9090) (22 (link)) (S1159), Novobiocin (NB) (23 (link)) (S2492), NVP-BEP800 (24 (link)) (S1498), MG132 (S2619), and cycloheximide (CHX) (S7418) were purchased from SelleckChem (USA).
Nvp bep800
Manufactured by Selleck Chemicals
Sourced in United States
The NVP-BEP800 is a laboratory equipment product. It is designed for use in research and testing environments. The core function of the NVP-BEP800 is to perform specific laboratory tasks, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Mg132 s2619, by Selleck Chemicals (1 mentions)
Ganetespib, by Selleck Chemicals (1 mentions)
Anti actin, by Merck Group (1 mentions)
Anti ripk1, by BD (1 mentions)
Anti vdac, by Merck Group (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Mabc604, by Merck Group (1 mentions)
Anti hsp90 adi spa 835, by Enzo Life Sciences (1 mentions)
Q vd oph, by R&D Systems (1 mentions)
17 aag, by Selleck Chemicals (1 mentions)
Varioskan flash microplate reader, by Thermo Fisher Scientific (1 mentions)
Ver 155008, by Selleck Chemicals (1 mentions)
4 protocols using nvp bep800
1
Cell Line Characterization and Compound Screening
2
Western Blot Analysis of Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary antibodies were used for western blots of both mouse and human cell lines: anti-MLKL (produced in-house;25 (link) available as MABC604, EMD Millipore, Billerica, MA, USA), anti-GAPDH (2118, Cell Signalling Technology, Danvers, MA, USA), anti-Actin (A-1987, Sigma-Aldrich, St Louis, MO, USA), anti-RIPK1 (610458, BD Biosciences, Franklin, NJ, USA), anti-Hsp90 (ADI-SPA-835, Enzo, Life Sciences, Farmingdale, NY, USA), anti-Cdc37 (D11A3, Cell Signalling Technology), and anti-VDAC (Sigma-Aldrich). Anti-mouse RIPK3 (PSC-2283-c100, Axxora, San Diego, CA, USA) and anti-human RIPK3 (ab56164, Abcam, Cambridge, UK) were used for their respective cell lines.
Recombinant hTNF-Fc, produced in-house, and the Smac mimetic, Compound A, were described previously.71 (link), 72 (link) QVD-OPh was obtained from R&D Systems (Minneapolis, MN, USA). Both NVP-BEP800 and 17-AAG were obtained from Selleckchem (Sydney, NSW, Australia), and AT13387 was produced by Active Biochem (Wanchai, Hong Kong).
Recombinant hTNF-Fc, produced in-house, and the Smac mimetic, Compound A, were described previously.71 (link), 72 (link) QVD-OPh was obtained from R&D Systems (Minneapolis, MN, USA). Both NVP-BEP800 and 17-AAG were obtained from Selleckchem (Sydney, NSW, Australia), and AT13387 was produced by Active Biochem (Wanchai, Hong Kong).
3
Regulation of VEGFR1/2 Expression by Hsp90
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs were seeded in 96-well plates. After 24 h, the plasmids, namely, pEZX–PG04–VEGFR1 and pEZX–PG04–VEGFR2 (FulenGene, China), were either transfected separately into cells, co-transfected with pcDNA3–Hsp90β or pRNAT-U6-siHsp90β, or incubated with NVP-BEP800 (Selleck, USA) for 48 h. Gaussia and secreted alkaline phosphatase (SEAP) luciferase activities were consecutively measured by a dual-luciferase reporter assay system (FulenGene, China) after 48 h of transfection. Gaussia luciferase activity was normalized to SEAP activity.
4
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed using MTT reagent Thiazolyl Blue Tetrazolium Bromide reconstituted in culture medium (5 mg/mL) to evaluate cell viability and proliferation, as has been described previously [69 (link)]. Briefly, cells were seeded at 100,000 cell/cm2 on 96-well culture plates and exposed to treatment conditions: BMSC-CM or ADSC-CM, recombinant proteins; HSPA5, HSPA9 or HSP90β1 (Aviva Systems Biology, San Diego, CA, USA) and HSP 70/90 family inhibitors; VER155008, HA15 or NVP-BEP800 (Selleckchem, Munich, Germany). After treatment, cells were washed with PBS, followed by incubation with MTT solvent for 3 h at 37 °C in a humidified incubator. Cell supernatant was replaced with dimethyl sulfoxide (DMSO) and absorbance readings were measured using the Varioskan™ Flash microplate reader (Thermo Scientific, Loughborough, UK) at 595 nm wavelength. The degree of cell viability was presented as a percentage of optical density (OD) relative to control.
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!