The human HCC cell line, Huh7, was purchased from the Riken Cell Bank (Tsukuba, Japan). DMEM (Life Technologies Japan Ltd., Tokyo, Japan) containing 10% FBS (Life Technologies Japan Ltd., Tokyo, Japan) was used to culture parental Huh7 cells. The sorafenib-resistant Huh7 cells were cultured in DMEM with 8.0μM sorafenib (sc-220125A, Santa Cruz, CA) dissolved in DMSO. The final concentration of DMSO was less than 0.1%.
Dulbecco modified eagle medium (dmem)
Manufactured by Santa Cruz Biotechnology
Sourced in Spain
DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium commonly used to support the growth and maintenance of a wide variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Sorafenib, by Santa Cruz Biotechnology (2 mentions)
Polybrene, by Santa Cruz Biotechnology (1 mentions)
Proteome profiler mouse xl cytokine array, by R&D Systems (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Rapamycin, by Merck Group (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Sirna transfection reagent, by Santa Cruz Biotechnology (1 mentions)
Penicillin streptomycin, by Santa Cruz Biotechnology (1 mentions)
Opti mem, by Santa Cruz Biotechnology (1 mentions)
P ampk, by Cell Signaling Technology (1 mentions)
Antibodies against p akt, by Cell Signaling Technology (1 mentions)
P mtor, by Cell Signaling Technology (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Compound c, by Merck Group (1 mentions)
8 bromo camp, by Merck Group (1 mentions)
Glucose assay kit, by Merck Group (1 mentions)
Fluorophore conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
9 protocols using dulbecco modified eagle medium (dmem)
1
Culturing Huh7 Hepatocellular Carcinoma Cells
2
Isolation and Characterization of Chondrocytes from R26Ikk2ca/Ikk2ca Mice
3
Autophagy Modulation Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3-MA, rapamycin, Chloroquine, compound C and Bafilomycin A1 were purchased from Sigma Aldrich (St. Louis, MO, USA). DMEM, Opti-MEM, penicillin-streptomycin, antibody against glyceraldehyde−3-phosphate dehydrogenase (GAPDH), AMPK siRNA, and siRNA Transfection Reagent were bought from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against LC3, p62 and LAMP2 were bought from Abcam (Cambridge, MA, USA), the antibodies against p-AKT, AKT, AMPK, p-AMPK, mTOR and p-mTOR were from Cell Signaling Technology (Danvers, MA, USA), the recombinant active full-length human Akt1 protein (rAkt1) was purchased from Abcam (Cambridge, MA, USA).
4
Glucose Production Assay in Hepatocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The glucose output assay was performed as previous reported.21 (link) Hepatocytes were washed 3 times with phosphate-buffered saline and were incubated in serum-free DMEM containing 0.5 mmol/L 8-bromo-cyclic adenosine monophosphate (cAMP) (Santa Cruz Biotechnology, Dallas, TX) for 5 hours. Cells then were incubated in 0.5 mL/well of phenol red–free, glucose-free DMEM (Sigma) containing 2 mmol/L pyruvate, 20 mmol/L lactate, and 1 mmol/L 8-bromo-cAMP, with 80 ng/mL leptin or vehicle. Medium was collected 3 hours later and subjected to glucose measurement using the Glucose Assay Kit (Sigma). Cells were lysed and the protein concentration was measured. The glucose production was normalized with cellular protein content.
5
HeLa Cell Culture Protocol
6
Protein Binding Assay in HEK293T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293T cells were grown in DMEM (Invitrogen) supplemented with 10% FBS (Invitrogen), penicillin/streptomycin, and transfected with EGFP, myc-LRRTM2, IgSF8-EGFP, and Tenascin-R-EGFP plasmids using Fugene6 (Promega). Twenty-four hours after transfection, the cells were incubated with Fc, NRXN1β-Fc, IgSF8-Fc, or Tenascin-R-Fc proteins (10 µg/ml) in DMEM supplemented with 20 mM Hepes (pH 7.4) for 1 h at RT. Following two brief washes in DMEM/20 mM Hepes pH 7.4, cells were fixed and immunostained using mouse anti-GFP (1:500; Santa Cruz), mouse anti-c-myc (1:1000; Santa Cruz), and the Cy3-conjugated anti-Fc antibody (1:1000; Jackson ImmunoResearch). Fluorophore-conjugated secondary antibodies were from Jackson ImmunoResearch or Invitrogen (used at 1:300 or 1:1000, respectively). The cells were imaged with a Leica SP8 confocal microscope (Leica Microsystems) using a 63× objective.
7
Cell Culture Protocol for Various Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BS-C-1 (ATCC), CV-1 (ATCC), MEFs (A gift from Prof. Dr Eugen Kerkhoff), HEK 293T (ATCC), A549 (ATCC), 2fTGH (Sigma-Aldrich), U3A (a 2fTGH derived STAT1-/- cell line, Sigma-Aldrich), U6A (a 2fTGH derived, STAT2-/- cell line, Sigma-Aldrich), and TK- 143B cells (ATCC) were maintained in DMEM (Gibco) supplemented with 10% (v/v) foetal bovine serum (FBS, PAN-Biotech) and penicillin-streptomycin (PS, 50 μg/mL, Gibco). T-REx 293 cells (Life technologies) were maintained in DMEM supplemented with 10% (v/v) FBS, PS (50 μg/mL) and blasticidin (10 μg/mL, Santa Cruz), and T-REx 293 derived cells lines expressing EV, TAP-N1, TAP-018, or TAP-NiV-V were further supplemented with zeocin (100 ug/mL, Invivogen). HeLa (ATCC) and RK13 cells (ATCC) were maintained in MEM (Gibco) supplemented with 10% (v/v) FBS, PS (50 μg/mL) and 1% (v/v) 100 X non-essential amino acids (Gibco). The construction of T-REx 293 cell lines expressing proteins inducibly is outlined in the method details section.
8
Activation of Cancer-Associated Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We purchased Huh‐7 (human HCC) cells from the Riken Cell Bank. The Huh7 cells were cultured in DMEM (Life Technologies Japan Ltd.) containing 10% FBS (Life Technologies Japan Ltd.). We developed the SR Huh7 (Huh7SR) cell line by culturing the cells in DMEM with 8.0 μM sorafenib (sc‐220125A; Santa Cruz Biotechnology), which was first dissolved in DMSO. The last concentration in DMSO was <0.1%. Lx2 human hepatic stellate cells were purchased from Merck Millipore (Tokyo, Japan). Huh7 and Huh7SR were cultured in DMEM for 48 h, then the cell‐culture medium was collected and centrifuged at 50g. Lx2 cells were co‐cultured with this conditioned medium or DMEM at a 1:1 ratio for at least 48 h (Figure 2A ). Activation of CAFs was confirmed using real‐time‐PCR (RT‐PCR) of α‐smooth muscle actin (α‐SMA) and fibroblast activation protein (FAP).11 (link)
9
661W Cell Line Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
661w cell line (a generous gift of Profs. N. Cuenca and V. Maneu) was grown in Dulbecco's modi ed Eagle medium (DMEM, SantaCruz Biotechnology) suplemented with 10% fetal bovine serum (Sigma-Aldrich, Madrid, Spain), 100 µg/ml penicillin and 100 u/ml streptomycin. Cultures were maintained in a controlled gas (95% air, 5% CO 2 ), humidity (95%) and temperature (37ºC) environment. Proliferation rate was assayed by allowing cells plated at several densities to grow for 1 to 3 days in 96-well plates. Then cells were incubated with resazurin that is reduced to uorescent resoru n by cell dehydrogenases. Fluorescence intensity is proportional to the number of cells. Doubling time was approximately 26h. This result was in line with a previous 661W growth rate estimation (Crawford et al., 2001) .
Appropriate number of cells were seeded in 75 cm 2 asks to reach con uency every 2-3 days when they were trypsinized, counted and plated in 96-well black with clear bottom plates for uorescence (Corning Incorporated, USA) at a density of 50,000 cells/well. All experiments were carried out 24h after plating.
Appropriate number of cells were seeded in 75 cm 2 asks to reach con uency every 2-3 days when they were trypsinized, counted and plated in 96-well black with clear bottom plates for uorescence (Corning Incorporated, USA) at a density of 50,000 cells/well. All experiments were carried out 24h after plating.
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!