Expi293F cells (Thermo) were maintained in Expi293 medium (Thermo) supplemented with 0.5% Penicillin-Streptomycin-Amphotericin B (Wako). HEK293S GnT1 -cells were maintained in FreeStyle293 medium supplemented with 0.5% FBS, 0.5% Penicillin-Streptomycin-Amphotericin B (Wako), 4 mM L-alanyl-L-glutamine (Wako). Cells used for protein expression were cultured in a Pro293s CDS medium (Lonza) supplemented with 0.5% FBS, 0.5% Penicillin-Streptomycin-Amphotericin B (Wako), 4 mM L-alanyl-L-glutamine (Wako).
Penicillin streptomycin amphotericin b
Manufactured by Fujifilm
Sourced in Japan, United States
Penicillin/streptomycin/amphotericin B is a combination of antibiotics and antifungal agents commonly used in cell culture applications. It serves as a broad-spectrum antimicrobial solution to help prevent bacterial and fungal contamination in cell culture media and samples.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Glutamax 1, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Hepg2, by RIKEN Cell Bank (2 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (2 mentions)
Expi293f cells, by Thermo Fisher Scientific (1 mentions)
Expi293 medium, by Thermo Fisher Scientific (1 mentions)
L alanyl l glutamine, by Fujifilm (1 mentions)
Huvecs, by Takara Bio (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Ficoll paque, by GE Healthcare (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Dmem with l glutamine and phenol red, by Fujifilm (1 mentions)
Hek293, by RIKEN Cell Bank (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
Mycoalert, by Lonza (1 mentions)
Snu 1, by American Type Culture Collection (1 mentions)
20 protocols using penicillin streptomycin amphotericin b
1
Protein Expression in Expi293F and HEK293S Cells
2
Culturing HUVECs and Mouse Primary SSCs
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HUVECs (TaKaRa Bio) were cultured using EGM2 (TaKaRa Bio) and penicillin/streptomycin/amphotericin B (Wako). Isolated mouse primary SSCs were cultured using the mouse MesenCult Expansion Kit with L-glutamine (Wako) and penicillin/streptomycin/amphotericin B (Wako).
3
Canine Bone Marrow-Derived Progenitor Cells
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Canine BM-PACs were cultured according to a previously described method [22 (link)] with modifications in the composition of the growth
medium after ceiling culture. Briefly, bone marrow aspirated from the proximal humerus was
subjected to density gradient centrifugation with Ficoll-Paque (GE Healthcare, Little
Chalfont, U.K.). After centrifugation, the top adipose layer containing mature adipocytes
was transferred into a new tube and washed twice with Dulbecco’s modified Eagle’s medium
(DMEM; Invitrogen, Carlsbad, CA, U.S.A.) supplemented with 10% fetal bovine serum (FBS;
Invitrogen) and 1% penicillin-streptomycin/amphotericin-B (Wako, Tokyo, Japan). The mature
adipocytes, to which BM-PACs adhered, were then placed in 25-cm2 flasks
completely filled with DMEM supplemented with 20% FBS and 1% antibiotics and subjected to
ceiling culture at 37°C in a humidified atmosphere containing 5% CO2. The
ceiling culture was maintained for 5–7 days without replacing the medium until the cells
reached 100% confluence. The confluent cells were detached with 0.25% trypsin/1 mM
ethylenediaminetetraacetic acid solution (Wako) and passaged at a density of 1 ×
104 cells/cm2 in growth medium consisting of DMEM, 10% FBS, 1%
antibiotics, and 10 ng/ml human recombinant basic
fibroblast growth factor (bFGF; Peprotech, Rocky Hill, NJ, U.S.A.).
medium after ceiling culture. Briefly, bone marrow aspirated from the proximal humerus was
subjected to density gradient centrifugation with Ficoll-Paque (GE Healthcare, Little
Chalfont, U.K.). After centrifugation, the top adipose layer containing mature adipocytes
was transferred into a new tube and washed twice with Dulbecco’s modified Eagle’s medium
(DMEM; Invitrogen, Carlsbad, CA, U.S.A.) supplemented with 10% fetal bovine serum (FBS;
Invitrogen) and 1% penicillin-streptomycin/amphotericin-B (Wako, Tokyo, Japan). The mature
adipocytes, to which BM-PACs adhered, were then placed in 25-cm2 flasks
completely filled with DMEM supplemented with 20% FBS and 1% antibiotics and subjected to
ceiling culture at 37°C in a humidified atmosphere containing 5% CO2. The
ceiling culture was maintained for 5–7 days without replacing the medium until the cells
reached 100% confluence. The confluent cells were detached with 0.25% trypsin/1 mM
ethylenediaminetetraacetic acid solution (Wako) and passaged at a density of 1 ×
104 cells/cm2 in growth medium consisting of DMEM, 10% FBS, 1%
antibiotics, and 10 ng/ml human recombinant basic
fibroblast growth factor (bFGF; Peprotech, Rocky Hill, NJ, U.S.A.).
4
Culturing Diverse Cell Lines for Research
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Human biliary cancer cells (NOZ), mouse embryonic fibroblasts (MEF) (Japanese Collection of Research Bioresources, Tokyo, Japan), human breast cancer cells (MCF7), human hepatocyte carcinoma cells (HepG2) and human embryonic kidney 293 cells (HEK293) (Riken Cell Bank, Ibaraki, Japan) were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) with L-glutamine and phenol red (Fujifilm Wako Pure Chemical Corp., Osaka, Japan) in a humidified atmosphere with 5% CO2 at 37°C. The medium was supplemented with 10% (v/v) heat-inactivated fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and penicillin-streptomycin-amphotericin B (Fujifilm Wako).
5
Culturing Colorectal Tumor Cell Lines
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The human colorectal tumor cell lines HCT116, DLD‐1, and RKO cells, gifted by Dr. Bert Vongelstein (Johns Hopkins University, Baltimore, MD, USA), as well as SW480 (EC87092801, ECACC, UK) and HT29 (EC91072201, ECACC) cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), 1% GlutaMAX‐I (Thermo Fisher Scientific), and 1% penicillin/streptomycin/amphotericin B (Wako Pure Chemical Industries, Osaka, Japan). The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2. Cells were harvested using 0.25% Trypsin–EDTA (Thermo Fisher Scientific, MA, USA) for further analysis.
6
CRC Cell Lines Cultivation Protocol
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Human CRC cell lines (DLD-1, HCT116, HT29, RKO, and SW480) were a gift from Dr. Bert Vogelstein (Johns Hopkins University, Baltimore, MD). The cells were incubated in Dulbecco’s modified Eagle’s medium (DMEM; Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific Inc., Waltham, MA), 1% GlutaMAX-I (Thermo Fisher Scientific Inc.), and 1% penicillin/streptomycin/amphotericin B (Wako Pure Chemical Industries Ltd., Osaka, Japan). The cells were maintained at 37 °C in a humidified atmosphere containing 5% CO2.
7
Gastric Cancer Cell Line Characterization
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The human gastric cancer cell lines MKN1 (RRID, CVCL_1415), MKN74 (CVCL_2791), MKN45 (CVCL_0434), and NUGC3 (CVCL_1612), as well as HeLa cervical carcinoma cells (CVCL_0030), were obtained from the JCRB cell bank. The human gastric cancer cell lines SNU1 (CVCL_0099), Hs746T (CVCL_0333), and NCI‐N87 (CVCL_1603) were from American Type Culture Collection and SNU216 (CVCL_3946) was from the Korean Cell Line Bank. The cells were maintained under a humidified atmosphere of 5% CO2 at 37°C in RPMI 1640 medium (Sigma‐Aldrich) supplemented with 10% heat‐inactivated fetal bovine serum (FBS) (Biowest) and 1% penicillin‐streptomycin‐amphotericin B (Wako). Cells were tested for mycoplasma contamination using MycoAlert (LT07, Lonza) and were confirmed negative. Eribulin was obtained from Eisai Co. Ltd and Oxaliplatin from Yakult. MORAb‐202 and farletuzumab were provided by Eisai Co. Ltd. MG132 was obtained from Funakoshi.
8
Cancer Cell Line Culture Protocols
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The esophageal cancer cell line KYSE30 (94072011; European Collection of Authenticated Cell Cultures, Salisbury, UK) was cultured in a 1:1 mixture of Ham’s F12 medium (Wako Pure Chemical Industries, Tokyo, Japan) and RPMI 1640 medium (Wako Pure Chemical Industries) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Biosera, Nuaillé, France) and 1% ampicillin and streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). The human colon cancer cell line HCT116 (CCL-247; American Type Culture Collection, Manassas, VA, USA) was cultured in RPMI 1640 medium (Wako Pure Chemical Industries) supplemented with 10% FBS and 1% ampicillin and streptomycin (Thermo Fisher Scientific). Murine colon adenocarcinoma cell line MC38 (ENH204-FP; Kerafast, Boston, MA, USA) was cultured in Dulbecco’s modified Eagle medium with high glucose (Wako Pure Chemical Industries) supplemented with 10% FBS, 1 mM sodium pyruvate (Thermo Fisher Scientific), 100 μM non-essential amino acids (Thermo Fisher Scientific), 50 μg/mL gentamicin (Wako Pure Chemical Industries), 10 μM 4-(2-hydroxyethyl)-1-piperazineëthanesulfonic acid (Thermo Fisher Scientific), and 1% penicillin-streptomycin-amphotericin B (Wako Pure Chemical Industries). Cells were cultured in an atmosphere of 5% carbon dioxide (CO2) at 37°C. All experiments using cells in this study were performed for fewer than 20 passages after thawing.
9
PBMC Activation and CD8+ T Cell Stimulation
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Suspension culture flasks (Sumitomo Bakelite, Tokyo, Japan) were pre-coated with 5 μg/ml anti-CD3 mAb and 5 μg/ml anti-CD28 mAb by incubation at room temperature for 45 min. PBMCs (1 × 105–1 × 106/ml) were suspended in AlyS505N-7 medium (Cell Science & Technology) containing 700 IU/ml IL-2, 5% FBS, and 1% penicillin-streptomycin-amphotericin B (Fujifilm Wako). The cells were then seeded into the flacks pre-coated with anti-CD3 and anti-CD28 mAbs. For short-term stimulation of CD8+ T cells derived from PBMCs, Dynabead human T-activator CD3/CD28 (Thermo Fisher Scientific) was used.
10
EGFR-Mutant Cell Line Characterization
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PC9Mock and PC9HRG cell lines have been previously characterized; these possess EGFR exon 19 deletions [22 (link)]. PC9HRGs were stably transfected with a full-length cDNA fragment encoding human heregulin (NRG1, NM_13956) as previously described [22 (link)]. Cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin-amphotericin B (Wako Pure Chemical Industries, Ltd., Osaka, Japan). We used cell lines within 6 months from resuscitation. Afatinib and erlotinib were obtained from commercial sources. Stock solutions of 10 mM afatinib and erlotinib were prepared in dimethylsulfoxide and stored at −20°C.
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