Human embryonic kidney (HEK) 293, HEK 293T, and human bone osteosarcoma epithelial (U2OS) cells were cultured as adherent cells directly on tissue culture plastic (Greiner) in Dulbecco’s modified Eagle’s medium (DMEM) growth medium (high glucose, l -glutamine, no sodium pyruvate; Gibco) with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) at 37°C and 5% CO2. All cells were harvested using 0.05% trypsin-EDTA (Gibco). Transfections were performed with TransIT-LT1 (Mirus). The panel of U2OS cells containing an integrated reporter (DR-GFP, SA-GFP, EJ2-GFP, and EJ5-GFP) used in the I-SceI repair assays were kindly provided by Jeremy M. Stark (Beckman Research Institute of the City of Hope) (60 ).
Tissue culture plastic
Manufactured by Greiner
Sourced in Germany
Tissue culture plastics are a type of laboratory equipment designed for the in vitro cultivation of cells, tissues, or organisms. These products provide a controlled and sterile environment for the growth and maintenance of biological samples. The core function of tissue culture plastics is to support the cultivation and study of various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Trypsin edta, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Mln7243, by Active Motif (3 mentions)
Dulbecco s modified, by Thermo Fisher Scientific (2 mentions)
Transit lt1, by Mirus Bio (2 mentions)
Dmem growth medium, by Thermo Fisher Scientific (2 mentions)
Brij r 35 solution, by Merck Group (2 mentions)
Terbium labeled streptavidin tb sa, by PerkinElmer (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Medium 199, by Lonza (1 mentions)
Foetal calf serum, by Thermo Fisher Scientific (1 mentions)
Endothelial cell growth supplement (ecgs), by PromoCell (1 mentions)
Trypsin edta, by Lonza (1 mentions)
Penicillin streptomycin pen strep, by Lonza (1 mentions)
Minimal essential medium, by Lonza (1 mentions)
Z vad fmk, by Peptide Institute (1 mentions)
Imatinib, by LC Laboratories (1 mentions)
Leibovitz s l 15 medium, by Thermo Fisher Scientific (1 mentions)
Eagle s minimum essential medium, by Merck Group (1 mentions)
9 protocols using tissue culture plastic
1
Culturing and Transfecting Human Cell Lines
2
Cell Culture Protocol for J-Lat, 5A8, and U2OS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The J-Lat 10.6 and 5A8 cells were cultured in flasks (Thermo Fisher Nunc™ EasYFlask™) in RPMI 1640 growth medium (L-glutamine, Gibco) with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) and the human bone osteosarcoma epithelial (U2OS) cells were cultured as adherent cells directly on tissue culture plastic (Greiner) in DMEM growth medium (high glucose, L-glutamine, no sodium pyruvate; Gibco) with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) at 37°C and 5% CO2. U2OS cells were harvested using 0.05% Trypsin-EDTA (Gibco).
3
Cell Culture and Stable Knockdown Generation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human bone osteosarcoma epithelial (U2OS), Human embryonic kidney (HEK) 293, and HEK 293T cells were cultured as adherent cells directly on tissue culture plastic (Greiner) in Dulbecco’s modified Eagle’s medium (DMEM) growth medium (high glucose, L-glutamine, no sodium pyruvate; Gibco) with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin-streptomycin (Gibco) at 37°C and 5% CO2. All cells were harvested using 0.05% trypsin-EDTA (Gibco). The panel of U2OS cells stably expressing 53PB1-GFP (64 (link)) and NBS1-GFP (65 (link)) were kindly provided by Claudia Lukas (University of Copenhagen, Denmark). Stable DCAF1 knock-down cells were generated by delivering lentiviruses expressing DCAF1 shRNAs or empty vector to U2OS cells. U2OS with integrated lentiviral construct were selected for with 1ug/ml of Puromycin. Knock-down of DCAF1 was assessed through western blots.
4
Culturing human cell lines for gene repair studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human embryonic kidney (HEK) 293, HEK 293T, and human bone osteosarcoma epithelial (U2OS) cells were cultured as adherent cells directly on tissue culture plastic (Greiner) in DMEM growth medium (high glucose, L-glutamine, no sodium pyruvate; Gibco) with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) at 37°C and 5% CO2. All cells were harvested using 0.05% Trypsin-EDTA (Gibco). Transfections were performed with TransIT-LT1 (Mirus). The panel of U2OS cells containing an integrated reporter (DR-GFP, SA-GFP, EJ2-GFP, EJ5-GFP) used in the I-SceI repair assays were kindly provided by Jeremy M. Stark (Beckman Research Institute of the City of Hope) (60) .
5
Endothelial Cell Culture Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Foetal calf serum (FCS) was purchased from Invitrogen Life Technologies, Scotland. Penicillin/streptomycin (Pen/Strep), trypsin/EDTA, minimal essential medium, α-modification (α-MEM) and Medium 199 and other tissue culture reagents were purchased from Lonza, Nottingham, UK. Endothelial cell growth supplement (ECGS) was obtained from Promocell, Heidelberg, Germany. Alkaline phosphatase staining reagents and assay kit and other cell culture reagents were purchased from Sigma Aldrich, Poole, Dorset, UK. Human VEGF-165 was purchased from PeproTech EC Ltd., London, UK. Collagenase B was purchased from Roche Diagnostic Ltd., Lewes, East Sussex, UK. Tissue culture plastics were purchased from Greiner BioOne, okGloucestershire, UK.
6
Binding Assay for IAP Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue culture plastics were purchased from Greiner Bio-One (Frickenhausen, Germany). The biotinylated anti-His antibody (PV6090) and the His-tagged ABL1 protein (full length, P3049) were from Life Technologies (Carlsbad, CA, USA). Brij(R) 35 solution was obtained from Merck Millipore (Billerica, MA, USA). Terbium-labeled streptavidin (Tb-SA) was from Cisbio (Codolet, France). The recombinant His-tagged human XIAP (BIR3, Asn252-Thr356, 895-XB-050) protein was purchased from R&D Systems (Minneapolis, MN, USA). Recombinant His-tagged human cIAP1 (BIR3, Leu250-Gly350) and cIAP2 (BIR3, Gln238-Ser349) proteins were expressed in E. coli and purified using a Ni-NTA column and gel filtration chromatography. The FITC-labeled Smac peptide (FITC-Smac, AVPIAQK(5-FAM)-NH2)56 (link) was synthesized in Scrum (Tokyo, Japan). The BODIPY-FL labeled dasatinib (BODIPY-dasatinib)57 (link) was synthesized as described previously. zVAD-FMK was purchased from Peptide Institute (Osaka, Japan).
7
In Vitro Binding Assays for IAP Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue culture plastics were purchased from Greiner Bio‐One (Tokyo, Japan). Imatinib was purchased from LC Laboratories (Woburn, MA, USA). MLN7243 was purchased from Active Biochem (Maplewood, NJ, USA). Cycloheximide (CHX) was from Sigma‐Aldrich (St. Louis, MO, USA). His‐tagged ABL1 protein (full length, P3049) was from Life Technologies (Carlsbad, CA, USA). Brij(R) 35 solution was obtained from Merck Millipore (Billerica, MA, USA). Terbium‐labeled streptavidin (Tb‐SA) was from Cisbio (Codolet, France). Recombinant His‐tagged human XIAP (BIR3, Asn252‐Thr356, 895‐XB‐050) protein was purchased from R&D Systems (Minneapolis, MN, USA). Recombinant His‐tagged human cIAP1 (BIR3, Leu250‐Gly350) and cIAP2 (BIR3, Gln238‐Ser349) proteins were expressed in E. coli and purified using a Ni‐NTA column and a gel filtration chromatography. FITC‐labeled Smac peptide (FITC‐Smac, AVPIAQK(5‐FAM)‐NH2)34 was synthesized in Scrum (Tokyo, Japan). BODIPY‐FL labeled dasatinib (BODIPY‐dasatinib)35 was synthesized as described previously.
8
Reagents for Cell Culture Experiments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue culture plastics were purchased from Greiner Bio‐One. RPMI 1640 medium, DMEM, Eagle's minimum essential medium, and kanamycin were from Merck. McCoy's 5a medium and Leibovitz's L‐15 medium were from Thermo Fisher Scientific. Benzyloxycarbonyl‐L‐leucyl‐L‐leucyl‐L‐leucinal (MG132) was from Peptide Institute. MLN7243 and MLN4924 were from Active Biochem.
9
Reagents for Cell Culture Experiments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue culture plastics were purchased from Greiner Bio-One (Frickenhausen, Germany).
Cycloheximide was purchased from Sigma-Aldrich (St. Louis, MO, USA). MG132 was purchased from Peptide Institute (Osaka, Japan). MLN7243 was purchased from Active Biochem (Maplewood, NJ, USA). Pomalidomide was purchased from Cayman Chemical (Ann Arbor, MI, USA). A23187 was purchased from Merck KGaA (Darmstadt, Germany).
Cycloheximide was purchased from Sigma-Aldrich (St. Louis, MO, USA). MG132 was purchased from Peptide Institute (Osaka, Japan). MLN7243 was purchased from Active Biochem (Maplewood, NJ, USA). Pomalidomide was purchased from Cayman Chemical (Ann Arbor, MI, USA). A23187 was purchased from Merck KGaA (Darmstadt, Germany).
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!