Human NF1-associated MPNST cell lines NF90.8 and ST88–14 were provided by Dr. Michael Tainsky (Wayne University, Detroit, MI) and sNF96.2 was purchased from ATCC (Manassas, VA). NF90–8, ST88–14, sNF96.2 and STS26T MPNST cell lines were cultured in DMEM (ATCC) media supplemented with 10% FBS (Sigma) and penicillin/streptomycin (Gibco). These cell lines were not authenticated. Human Schwann cells isolated from spinal nerve were purchased from ScienCell Research Laboratories and maintained in DMEM media supplemented with 3% FBS, penicillin/streptomycin, and heregulin-1 (PeproTech) 20 ng/ml and 2 μM forskolin (Sigma). All cells were tested and found free of mycoplasma contamination.
Snf96
Manufactured by American Type Culture Collection
Sourced in United States, United Kingdom
The SNF96.2 is a laboratory equipment product offered by American Type Culture Collection. It serves as a centrifuge designed to separate and isolate biological materials, such as cells or cellular components, based on their density differences when subjected to centrifugal force. The core function of the SNF96.2 is to facilitate the purification and analysis of various biological samples through the process of sedimentation.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions)
Snf02, by American Type Culture Collection (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Forskolin, by Merck Group (4 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Heregulin 1, by Thermo Fisher Scientific (2 mentions)
Snf94, by American Type Culture Collection (2 mentions)
Rapamycin, by Merck Group (1 mentions)
Anti p62, by Cell Signaling Technology (1 mentions)
Anti atg7, by Cell Signaling Technology (1 mentions)
Anti lc3, by Cell Signaling Technology (1 mentions)
Anti bax, by Cell Signaling Technology (1 mentions)
Lbh589, by Selleck Chemicals (1 mentions)
Anti bcl 2, by Cell Signaling Technology (1 mentions)
Anti cyclin d1, by Cell Signaling Technology (1 mentions)
Anti beclin 1, by Cell Signaling Technology (1 mentions)
Anti atg12, by Cell Signaling Technology (1 mentions)
19 protocols using snf96
1
NF1-associated MPNST Cell Line Protocols
2
NF1-associated MPNST Cell Line Protocols
3
Cell Culture and Characterization of MPNST
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human MPNST cell lines sNF96.2 and sNF02.2 were purchased from ATCC (ATCC, Manassas, VA), while ST8814 and STS26T cells were kind gifts from Dr. Yang Jilong (Tianjin Medical University, China) and Dr. Nancy (Cincinnati Children’s Hospital Medical Center, USA). All cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% FBS at 37 °C in a humidified atmosphere with 5% CO2. Neurofibroma samples were obtained from NF1 patients after informed consent. Tumour specimens were obtained from 3 different patients (2 males and 1 female; age 20–47 years). Separation and culture of neurofibroma Schwann cells (NFSCs) were performed according to the protocol by Thorsten Rosenbaum [24 (link)].
Rapamycin and 3-methyladenine (3MA) were purchased from Sigma Chemical Co. LBH589 was purchased from Selleckchem. The following antibodies were used in the experiments: anti-HMGA2, anti-cyclin D1, anti-BCL2, anti-Bax, anti-LC3, anti-p62, anti-Beclin1, anti-PARP, anti-PTEN, anti-ATG12, anti-ATG7,anti-acetyl histone 3 (H3) antibody and anti-GAPDH antibodies from Cell Signaling Technology (Beverly, MA, USA); anti-MSI2 antibodies from Abcam (Cambridge, MA, USA) and Novus Biologicals; and anti-NF1 antibodies from Bethyl (Montgomery, TX, USA).
Rapamycin and 3-methyladenine (3MA) were purchased from Sigma Chemical Co. LBH589 was purchased from Selleckchem. The following antibodies were used in the experiments: anti-HMGA2, anti-cyclin D1, anti-BCL2, anti-Bax, anti-LC3, anti-p62, anti-Beclin1, anti-PARP, anti-PTEN, anti-ATG12, anti-ATG7,anti-acetyl histone 3 (H3) antibody and anti-GAPDH antibodies from Cell Signaling Technology (Beverly, MA, USA); anti-MSI2 antibodies from Abcam (Cambridge, MA, USA) and Novus Biologicals; and anti-NF1 antibodies from Bethyl (Montgomery, TX, USA).
4
Melanoma Cell Line Cultivation Protocol
5
Characterization of MPNST Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One HSC line and two MPNST cell lines (sNF02.2, sNF96.2) were purchased from ATCC (American Type Culture Collection, ATCC). Five MPNST cell lines (ST88-14, STS26T, T265, S462, and S462TY) were kindly granted by Prof. Vincent Keng (Li X.X. et al., 2018 (link)) and Prof. Jilong Yang (Du et al., 2013 (link)). All MPNST cell lines were derived from NF1 patients, except STS26T. Cell lines were maintained in DMEM, 10% FBS and penicillin/streptomycin (Gibco, United States) and were tested mycoplasma negative every 3 months. Verification of cell lines was confirmed by Short Tandem Repeat DNA profiling (Applied Biological Materials Inc., Canada).
6
Schwann Cell Culture and ILB Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human Schwann cells (sNF96.2, ATCC CRL-2884™, ATCC, Teddington, UK) were cultured in 25 cm2 culture flasks coated with poly-d-lysine and laminin) in high-glucose DMEM with 10% foetal bovine serum (FBS) (n = 8) and incubated at 37 °C/5%CO2. Twenty-four hours after seeding (Day 0) cells were treated once with either ILB® (0.01 mg/ml in DMEM) (n = 3) or culture media (n = 3). The remaining 2 flasks of Schwann cells were used as the baseline cultures. After 48 h with treatment, the media were removed and cells were harvested in 2.5 ml Trizol:Water (4:1) solution containing 0.5 ml of chloroform at room temperature (RT). The flasks were inspected under the microscope to ensure full removal of cells. The collected lysates were stored at −80 °C.
7
Malignant Peripheral Nerve Sheath Tumor Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human MPNST cell lines sNF96.2 and sNF02.2 were purchased from ATCC (ATCC, Manassas, VA), while ST8814 and STS26T cells were kind gifts from Dr. Yang Jilong (Tianjin Medical University, China) and Dr. Nancy (Cincinnati Children’s Hospital Medical Center, USA). All cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% FBS, and they were maintained at 37 °C in a humidified atmosphere with 5% CO2.
The following antibodies were used in the experiments: anti-E-cad, anti-N-cad, anti-Vimentin and anti-GAPDH antibodies from Cell Signaling Technology (Beverly, MA, USA); anti-MSI2 and anti-CAV1 antibodies from Abcam (Cambridge, MA, USA),anti-ubiquitin (FK2) from Enzo Life Sciences(New York, NY, USA).
The following antibodies were used in the experiments: anti-E-cad, anti-N-cad, anti-Vimentin and anti-GAPDH antibodies from Cell Signaling Technology (Beverly, MA, USA); anti-MSI2 and anti-CAV1 antibodies from Abcam (Cambridge, MA, USA),anti-ubiquitin (FK2) from Enzo Life Sciences(New York, NY, USA).
8
Immortalized Cells for NF1-Associated MPNST
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immortalized SCs (hTERT NF1 ipn02.3 2λ, M. Wallace), NF cells [ipn95.11bC, American Type Culture Collection (ATCC): Neurofibromatosis Therapeutic Acceleration Program (NTAP)], NF1-associated MPNST cells (sNF96.2, ATCC; MPNST642, MD Anderson; T265, A. Harder), and sporadic MPNST cells (STS26T, D. Scoles) were seeded on poly-l -lysine–coated 12-mm circle glass coverslips (Carolina Biologicals) at 50,000 cells in 24-well cell culture dishes. Coverslips were fixed in 4% (w/v) paraformaldehyde (PFA) for 20 min and washed in 1× phosphate-buffered saline (PBS) four times before immunofluorescence staining. For BrdU pulse labeling, cells were incubated with 20 mM BrdU for 4 hours before fixation.
9
Lentiviral Silencing of PKC Isoforms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human Nf1 deficient ST, SNF96.2 or proficient SNF02.2 cells were purchased from ATCC (Manassas, VA). The cells were cultured in Dulbecco's Modified Eagles's medium supplemented with 10% heat-inactivated Fetal Bovine Serum (Atlanta Biologicals), 100 units/ml penicillin, 100μg/ml Streptomycin (Invitrogen). ST cells were stably transfected with the Nf1 effective domain gene and maintained in the medium containing 400 μg/ml of G418 (Fisher Scientific Inc. MA). HMG was purchased from EMD Millipore (Billerica, MA). Antibodies were purchased from BD (San Jose, CA).
The oligonucleotides containing small interference RNA sequences targeting different Protein Kinase C isozymes were ligated to a lentiviral small-hairpin (sh) RNA expression vector pLentiLox3.7. The sequences of the shRNA α and β are: 5′-ggctgtacttcgtcatgga-3′ and 5′-caggaagtcatcaggaata-3′ for human PKC α and PKC β, respectively. FuGene 6 transfection reagent (Roche Applied Science, IN) was used for transfections.
The oligonucleotides containing small interference RNA sequences targeting different Protein Kinase C isozymes were ligated to a lentiviral small-hairpin (sh) RNA expression vector pLentiLox3.7. The sequences of the shRNA α and β are: 5′-ggctgtacttcgtcatgga-3′ and 5′-caggaagtcatcaggaata-3′ for human PKC α and PKC β, respectively. FuGene 6 transfection reagent (Roche Applied Science, IN) was used for transfections.
10
Cell Culture Protocols for Pancreatic Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All cell lines were maintained at 37 °C with 5% CO2. Panc-1, RSC96, sNF96.2, CAFs, hPSCs, and NF cells were cultured in DMEM (Meilunbio, China) containing 10% fetal bovine serum (FBS) (Gibco; Life Technology) and 50 μg/mL penicillin/streptomycin (P/S). CFPAC-1 cells were cultured in IMDM (BIOAGRIO) containing 10%FBS and 50 μg/mL P/S. Panc-1 (CRL-1469™), CFPAC-1 (CRL-1918™), RSC96 (CRL-2765™), and sNF96.2(CRL-2884™) were purchased from ATCC. NF and CAFs were isolated from human normal/tumor tissues. hPSC was a gift from Dr. Yuan Fang, which were purchased from ScienCell Research Laboratories, Carlsbad, CA (the HPaSteC cells, #3830). All cells were checked routinely for the absence of mycoplasma contamination. Short tandem repeat profiling was used to authenticate all the cell lines.
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!