H9c2, a rat cardiac myoblast cell line, was obtained from American Type Culture Collection (ATCC, Cat# CRL-1446, RRID: CVCL_0286). Dulbecco Minimal Essential Medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Cat# 11965-092, 10099-141). 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was obtained from Sigma-Aldrich (Cat# M2128). Ang II was purchased from Tocris Bioscience (Cat# 1158). Ghrelin was obtained from Phoenix Biotech (Cat# 031-31). The Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit and BCA Protein Quantitation Assay were purchased from KeyGen Biotech (Cat# KGA108, KGP902). PrimeScript™ RT reagent kit and SYBR fluorescent quantitation kit were provided by Takara (Cat# RR047A, RR420A). The Rat Apoptosis RT2 Profiler™ PCR Array and other related reagents were obtained from QIAGEN Bioinformatics (Cat# PARN-012Z). The Fugene transfection reagent and Dual-Luciferase Reporter Assay System were purchased from Promega (Cat# E2311, E1910). Primary antibodies against SESN2 (Cat# ab178518), β-ACTIN (Cat# ab16039), and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG secondary antibody (Cat# ab6721) were purchased from Abcam and the details of these antibodies are described in Table 1 .
Dulbecco minimal essential medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
Dulbecco's Modified Eagle's Medium (DMEM) is a cell culture medium commonly used for the maintenance and growth of various cell lines. It provides a balanced salt solution and essential nutrients required for cell proliferation. DMEM supports the culture of a wide range of mammalian cell types, including fibroblasts, epithelial cells, and many transformed cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin, by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (3 mentions)
Primescript rt reagent kit, by Takara Bio (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fetal calf serum fbs, by Thermo Fisher Scientific (2 mentions)
Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit, by Keygen Biotech (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (1 mentions)
Fugene transfection reagent, by Promega (1 mentions)
Ang 2, by Bio-Techne (1 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit igg secondary antibody, by Abcam (1 mentions)
Rat apoptosis rt2 profiler pcr array, by Qiagen (1 mentions)
Bca protein quantitation assay, by Keygen Biotech (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
A549 cell line, by American Type Culture Collection (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
8 protocols using dulbecco minimal essential medium
1
Cardiac Myoblast Cell Line H9c2 Assays
2
Culturing Lung Cancer and Control Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lung adenocarcinoma A549 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, USA), while normal human embryonic lung fibroblast HELF cell lines were purchased from cell bank (Chinese Academy of Sciences, Shanghai, China). Human non-small cell lung cancer A549 cells were cultured in RPMI-1640 medium (Gibco), containing 10% FBS, 100 mg/mL streptomycin (Invitrogen), 100 U/mL penicillin (Invitrogen), and maintained at 37°C with 5% CO2 at a humidified atmosphere. And human embryonic lung fibroblast HELF cells, used as corresponding control group, were cultured in Dulbecco minimal essential medium (Gibco) containing the same materials.
3
MYC Silencing in Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phosphate buffered saline (PBS), Dulbecco minimal essential medium (DMEM), Roswell Park Memorial Institute (RPMI)-1640 and fetal bovine serum (FBS) were purchased from Gibco (Thermo, UT, USA). Short interfering RNAs (siRNAs) targeting MYC were purchased from GenePharma (Shanghai, China). Lipofectamine 2000 and TRIzol were purchased from Invitrogen (Carlsbad, CA, USA). PrimeScriptTM RT reagent KIT and TB GreenTM Premix Ex TaqTM Ⅱ were from Takara (Otsu, Shiga, Japan). The CCK-8 proliferation kit was purchased from Dojindo Laboratories (Kumamoto, Japan). RIPA (Radio Immunoprecipitation Assay) Lysis Buffer, Bicinchoninic acid (BCA) protein assay kit, and Propidium Iodide (PI) kit were supplied by Beyotime Biotechnology (Shanghai, China).
4
Cell Culture and Mosquito Maintenance Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vero cells (African green monkey kidney) were cultured in serum free Opti-Pro medium (Invitrogen) as previously described [42 (link)]. Mosquito C6/36 (derived from A. albopictus; ATCC), C710 and Aag2 (derived from A. albopictus and A. aegypti, respectively; generous gift from Dr. A. Fallon, University of Minnesota) cells were maintained in Dulbecco minimal essential medium (DMEM) (Invitrogen) supplemented with 5% FBS (Lonza), 1x penicillin-streptomycin-glutamine (PSG) solution (Invitrogen), 1x MEM nonessential amino acids (Cellgro, Swedesboro, NJ), 1 × MEM vitamin solution (Invitrogen) and 5 μg/L of gentamicin (Invitrogen) at 32°C in an atmosphere of 5% CO2. The C6/36 cells were used in all but the initial experiments, because they exhibit superior viability in our experimental conditions compared to C710 cells.
Galveston colony of A. albopictus (generous gift of Dr. S. C. Weaver, UTMB) and NIH strain of A. aegypti mosquitoes have been described earlier [53 (link),68 (link)]. Both mosquito species were maintained in carton containers supplemented with 10% sucrose on cotton balls at 28°C, 80% humidity and with a 16-hr daylight cycle.
Galveston colony of A. albopictus (generous gift of Dr. S. C. Weaver, UTMB) and NIH strain of A. aegypti mosquitoes have been described earlier [53 (link),68 (link)]. Both mosquito species were maintained in carton containers supplemented with 10% sucrose on cotton balls at 28°C, 80% humidity and with a 16-hr daylight cycle.
5
Evaluation of Cytotoxic Compounds in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Aconitine, Hypaconitine, Mesaconitine, Higenamine, Hypaconine, Mesaconine, Talatisamine, Glycyrrhizic acid, Quercetin and 6-Gingerol (purity 99%) were purchased from Shanghai standard Corporation (http://naturestandard.cn.alibaba.com ). The structures of above chemicals were unambiguously identified by 1H NMR and MS spectra, and their purity were over 99% determined by HPLC-UV. Recombinant mouse TNF-α were purchased from Sigma (St.Louis, MO, USA). Actinomycin D was purchased from Sigma (St.Louis, MO, USA). Necrostatin-1 was purchased from Selleck Chemicals (Houston, USA). L929 mouse fibroblast cell line and Rat cardiac H9C2 cell line were obtained from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Dulbecco minimal essential medium (DMEM) was purchased from Invitrogen Corporation (Grand Island, NE, USA) and supplemented with 10% fetal calf serum (FBS) obtained from Gibco Co. (Rockville, MD, USA). Dimethyl sulfoxide (DMSO), penicillin streptomycin, and trypsin were purchased from Gibco Co. All experiments were repeated three times.
6
Cytosolic Ca2+ Response to OT and OT-GKR Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect potential difference in cytosolic Ca2+ concentration upon the treatment with OT and OT-GKR the experiments were performed on rat cardiac H9c2 cell line obtained from American Type Culture Collection (Rockville, MD, USA). Cells were grown on Dulbecco minimal essential medium (DMEM) purchased from Invitrogen Corporation (Grand Island, NE, USA) and supplemented with 10% fetal calf serum (FBS) from Gibco Co. (Rockville, MD, USA). Dimethyl sulfoxide (DMSO), penicillin streptomycin, and trypsin were purchased from Gibco Co. We analyzed the calcium response of cells in confluency of ∼300 cells/mm2. Changes in cytosolic Ca2+ concentration ([Ca2+]i) were monitored with radiometric Fura-2 fluorescence as described previously [17 (link)]. Fluorescence images were recorded at 15-sec intervals with a MicroMAX digital camera (Princeton Instruments Inc., Trenton, NJ). As a positive control for Ca2+ stimulation, 10 μM ionomycin (Invitrogen (Grand Island, NE), a Ca2+ ionophore (Sigma-Aldrich, St. Louis, MO) or 1 μM ATP (Sigma-Aldrich) was used. Changes of [Ca2+]i are presented as Fura-2 fluorescence F340/F380 ratio normalized to that value recorded in cells before stimulation.
7
Purification and Characterization of Talatizamine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Doxorubicin
hydrochloride,
tetracycline, dexamethasone, oxymetazoline, tamsulosin, benzoylaconitine,
and 4-aminopyridine (4-AP) were purchased from the National Institute
for the Pharmaceutical and Biological Products of China (Beijing,
China), and the purities were all over 98%. 25 mg of talatizamine
(TALA) was isolated and purified from the roots of A. carmichaeli by the authors. The structure of TALA was unambiguously identified
by 1H NMR and MS spectra, and its purity was over 98% determined
by HPLC-UV. A. carmichaeli (collection in Sichuan,
China) were purchased from Shanghai Leiyunshang Medicine Corp. (Shanghai,
China). Rat cardiac H9c2 cell line was obtained from American Type
Culture Collection (Rockville, MD, USA). Dulbecco minimal essential
medium (DMEM) was purchased from Invitrogen Corporation (Grand Island,
NE, USA) and supplemented with 10% fetal calf serum (FBS) obtained
from Gibco Co. (Rockville, MD, USA). Dimethyl sulfoxide (DMSO), penicillin
streptomycin, and trypsin were purchased from Gibco Co. The silica
gel (5 μm, 200 Å) was obtained from Meigao Materials, Inc.
(Qingdao, China). HPLC-grade acetonitrile was purchased from Merck
Co. (Darmstadt, Germany). MS-grade ammonia acetate was obtained from
Sigma Co. (St. Louis, MO, USA). Ultrapure water was prepared with
a Milli-Q water purification system (Millipore, Bedford, MA, USA).
Other reagents were of analytical grade.
hydrochloride,
tetracycline, dexamethasone, oxymetazoline, tamsulosin, benzoylaconitine,
and 4-aminopyridine (4-AP) were purchased from the National Institute
for the Pharmaceutical and Biological Products of China (Beijing,
China), and the purities were all over 98%. 25 mg of talatizamine
(TALA) was isolated and purified from the roots of A. carmichaeli by the authors. The structure of TALA was unambiguously identified
by 1H NMR and MS spectra, and its purity was over 98% determined
by HPLC-UV. A. carmichaeli (collection in Sichuan,
China) were purchased from Shanghai Leiyunshang Medicine Corp. (Shanghai,
China). Rat cardiac H9c2 cell line was obtained from American Type
Culture Collection (Rockville, MD, USA). Dulbecco minimal essential
medium (DMEM) was purchased from Invitrogen Corporation (Grand Island,
NE, USA) and supplemented with 10% fetal calf serum (FBS) obtained
from Gibco Co. (Rockville, MD, USA). Dimethyl sulfoxide (DMSO), penicillin
streptomycin, and trypsin were purchased from Gibco Co. The silica
gel (5 μm, 200 Å) was obtained from Meigao Materials, Inc.
(Qingdao, China). HPLC-grade acetonitrile was purchased from Merck
Co. (Darmstadt, Germany). MS-grade ammonia acetate was obtained from
Sigma Co. (St. Louis, MO, USA). Ultrapure water was prepared with
a Milli-Q water purification system (Millipore, Bedford, MA, USA).
Other reagents were of analytical grade.
8
Derivation and Cryopreservation of Mouse Embryonic Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse embryonic fibroblasts (MEFs) were collected from 13.5-day-old fetuses of outbred Institute of Cancer Research (ICR) mice. The visceral organs, head and extremities of the fetuses were removed and the remaining tissue was cut into small pieces. The manipulated tissue were subsequently incubated in 0.04% (v/v) trypsin-ethylenediaminetetraacetic acid (EDTA) (Gibco Invitrogen, Carlsbad, CA, USA) for 6 min with agitation, and centrifuged at 110×g for 2 min. The supernatants were diluted in 10% (v/v) fetal bovine serum (FBS; Hyclone Laboratories, Logan, UT, USA) containing Dulbecco minimal essential medium (DMEM; Gibco Invitrogen, USA). After being centrifuged at 390×g for 4 min one more time, the pellets were seeded into plastic culture dishes containing DMEM for monolayer formation. When the fibroblasts formed a confluent monolayer, they were cryopreserved using 10% dimethylsulfoxide (DMSO; Sigma Aldrich, St. Louis, MO, USA). After thawing, MEFs were treated with 10 μg/mL mitomycin C (Sigma Aldrich, USA) for 3 h and subsequently seeding in four-well multi-dishes (SPL, Pocheon, Korea)
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!