Mouse embryonic fibroblasts were harvested from GD15 embryos as previously described (Lipinski et al., 2006 (link)). Cells were grown to confluence in Dulbecco's modified Eagle's medium (DMEM) (with l-glutamine, 4.5 g l−1 glucose without sodium pyruvate) with 10% FCS and 1% penicillin-streptomycin. They were then trypsinized and plated in 24- or 48-well tissue culture plates (Falcon, Franklin Lakes, NJ) at 4.0×105 cells ml−1 media. Cells were allowed to attach for 24 h, and media were replaced with DMEM containing 1% FBS with or without the active SHH N-terminal peptide (R&D Systems, Minneapolis, MN) at 0.4 μg ml−1, with or without vismodegib (LC Laboratories) at indicated concentrations. For cell culture experiments, vismodegib was dissolved in DMSO.
48 well tissue culture plate
Manufactured by BD
Sourced in United States
48-well tissue culture plates are a type of laboratory equipment used for cell culture and tissue growth experiments. They provide a standardized format with 48 individual wells for culturing cells or tissues in a controlled environment.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Biowest (3 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Hepg2 cells, by RIKEN Cell Bank (2 mentions)
Shh n terminal peptide, by R&D Systems (1 mentions)
Vismodegib, by LC Laboratories (1 mentions)
Mouse gene 1.0 st array, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Expression console 1, by Thermo Fisher Scientific (1 mentions)
Mem neaa, by Thermo Fisher Scientific (1 mentions)
24 well tissue culture plate, by BD (1 mentions)
96 well tissue culture plate, by BD (1 mentions)
Dulbecco s modified eagle s medium, by Fujifilm (1 mentions)
8 protocols using 48 well tissue culture plate
1
Mouse Embryonic Fibroblast Culture and SHH Signaling
2
Sca-1+/CD34+/lin- Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sca-1+/CD34+/lin- cells were stained and sorted as mentioned above. Cells were plated at a density of 10,000 cells per well in 48 well tissue culture plates (BD Falcon) and cultured in α-MEM supplemented with 2mM Glutamax, 10% FBS, and penicillin/streptomycin at 37°C in 5% CO2. This medium was considered to be non-induction medium. Medium was changed every 3 days. At 7 days, cells were processed for RNA as mentioned above in the quantitative PCR section.
3
Culturing Caco-2 and HT-29 Colorectal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell lines used in this study were human colorectal adenocarcinoma Caco-2 (cancer coli-2) and colorectal adenocarcinoma cells HT-29. All cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with the following: (1) 10% fetal bovine serum, (2) antibiotics (100 U/mL penicillin and 100 mL/mL streptomycin), and (3) 25 μg/mL amphotericin B at 37 °C in an atmosphere of 5% CO2 (Eppendorf, Hamburg, Germany). Cells were incubated inside a 5% CO2 incubator at 37 °C for 5 days until they reached 90% confluence. Thereafter, we used PBS buffer to wash cells 2–3 times to get rid of the culture media or non-adherent cells. Before each assay, cells were seeded in 48-well tissue-culture plates (BD Falcon, Becton, NJ, USA) and incubated for 24 h inside a 5% CO2 incubator at 37 °C (at a density of 50,000 cells per well).
4
Microarray Analysis of Unstimulated Dendritic Cells
5
Fabrication of NO-Releasing PA Nanomatrices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As documented in previous work, a mixture of PA-YIGSR and PA-KKKKK at a 9:1 molar ratio was reacted with NO gas to generate PA-YK-NO.11 (link) In order to make NO-releasing PA nanomatrices, 120 μL of a 0.1% wt stock PA-YK-NO solution (pH 7.4) was prepared and dispensed into 48-well tissue culture plates (BD Biosciences, San Jose, CA, USA) with serial dilutions to make different PA-YK-NOs, ie, 130 μmol PA-YK-NO, 65 μmol PA-YK-NO, 32.5 μmol PA-YK-NO, and 16.25 μmol PA-YK-NO.
6
Cell Culture Conditions for HepG2 and HEK293
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2 and HEK293 cells were obtained from Riken cell bank (Tsukuba, Japan). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; WAKO Pure Chemicals, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS; Biowest, Miami, FL), minimum essential medium nonessential amino acids (MEM NEAA; Invitrogen, Carlsbad, CA), and Antibiotic-Antimycotic (Invitrogen), under 5% CO2 at 37°C. Before 24 h adenovirus infection, HepG2 cells were seeded at 1.5 × 10 5 cells per well onto 24-well tissue culture plates (BD Biosciences, Heidelberg, Germany) for real-time PCR, 1.0 × 10 5 cells per well onto 48-well tissue culture plates (BD Biosciences) for immunoblotting, and 0.5 × 10 5 cells per well onto 96-well tissue culture plates (BD Biosciences) for P450-Glo TM assay. The cells were subjected to corresponding experiments after 48 h.
7
HepG2 Cells Expressing Recombinant CYP P450s
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2 cells were purchased from RIKEN cell Bank (Tsukuba, Japan) and were cultured in Dulbecco’s modified Eagle’s medium (Wako Pure Chemical Industries) supplemented with 10% fetal bovine serum (Biowest, Miami, FL, USA), non-essential amino acids (Thermo Fisher Scientific, Maltham, MA, USA), and antibiotic-antimycotic (Thermo Fisher Scientific) under 5% CO2-95% air at 37 °C. The cells were seeded in a 48-well tissue culture plate (Becton Dickinson) at 5.0 × 104 cells/well. After 48 h, the cells were simultaneously infected with Ad-CYP1A2 at 5 multiplicity of infection (MOI), Ad-CYP2C9 at 1 MOI, Ad-CYP2C19 at 2 MOI, Ad-CYP2D6 at 0.05 MOI, and Ad-CYP3A4 at 10 MOI. The cells were cultured for 72 h and subsequently used as Ad-P450 cells. The cells were incubated in culture medium containing P450 substrate cocktail for 5 h.
8
Hepatocyte Cytochrome P450 Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2 cells were purchased from RIKEN Cell Bank (Tsukuba, Japan) and were cultured in DMEM (Wako Pure Chemical Industries), containing 10% fetal bovine serum (Biowest, Miami, FL, USA), nonessential amino acids (Thermo Fisher Scientific, Waltham, MA, USA), and Antibiotic-Antimycotic (Thermo Fisher Scientific) under 5% CO2 and 95% air at 37°C. The cells were seeded in a 48-well tissue culture plate (Becton Dickinson) at 5.0 × 10 4 cells/well. After 48 h, the cells were simultaneously infected with CYP1A2-expressing adenoviruses (Ad-CYP1A2) at five multiplicity of infections (MOIs): Ad-CYP2C9 at 1 MOI, Ad-CYP2C19 at 2 MOI, Ad-CYP2D6 at 0.05 MOI, and Ad-CYP3A4 at 10 MOI. The Ad-P450 cells were cultured for 72 h, incubated in culture medium containing each pharmaceutical (10 μmol/L) for 5 h, and subsequently extracted with ethyl acetate (1 mL). After evaporation of the organic layer, the residue was dissolved in the mobile phase, and an aliquot (0.4 μL) of the solution was subjected to LC-MS/MS.
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!