Six human gastric carcinoma cell lines, H-111-TC, HGC-27, Kato-III, MKN-1, MKN-45 and NU-GC-4 were purchased from Riken Cell Bank (Tsukuba, Japan) and used in the present study. According to the description by the Riken Cell Bank and literature (22 (link)–27 (link)), these cell lines originated from 1 unknown origin (H-111-TC) and 5 metastatic carcinoma, as well as 2 tubular (well differentiated or adenosquamous) carcinoma (H-111-TC and MKN-1) and 4 poorly differentiated carcinoma including signet ring cell type. HGC-27 and MKN-1 cell lines were maintained in Dulbeccos modified Eagles medium (DMEM; Gibco-BRL, Rockville, MD, USA), whereas NU-GC-4, Kato-III, MKN-45 and H-111-TC cell lines were maintained in RPMI-1640 (Gibco-BRL), supplemented with 10% fetal calf serum (FCS), penicillin, and streptomycin, at 37°C in a humidified atmosphere of 5% CO2. When they reached 80–90% confluence, the cells were washed with phosphate-buffered saline (PBS) and homogenized immediately in Isogen reagent (Nippon Gene, Osaka, Japan). Total RNA was extracted according to the manufacturers instructions.
Nugc4
Manufactured by RIKEN Cell Bank
Sourced in Japan
NUGC4 is a human gastric cancer cell line derived from a metastatic lymph node. It can be used for various cell culture-based applications.
Automatically generated - may contain errors
Lab products found in correlation
Mkn45, by RIKEN Cell Bank (6 mentions)
Hgc27, by RIKEN Cell Bank (5 mentions)
Mkn74, by RIKEN Cell Bank (5 mentions)
Kato 3, by RIKEN Cell Bank (3 mentions)
Rpmi 1640, by Nacalai Tesque (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Mkn45, by Thermo Fisher Scientific (1 mentions)
Hgc 27, by Thermo Fisher Scientific (1 mentions)
Kato 3, by Thermo Fisher Scientific (1 mentions)
Nu gc 4, by Thermo Fisher Scientific (1 mentions)
Isogen reagent, by Nippon Gene (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
1640 medium, by Merck Group (1 mentions)
Dulbecco s modified eagle medium, by Nacalai Tesque (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Ab17147, by Abcam (1 mentions)
Ab120952, by Abcam (1 mentions)
Anti β actin actb antibody, by Merck Group (1 mentions)
Pe anti pd l1 antibody, by BioLegend (1 mentions)
Ab157489, by Abcam (1 mentions)
6 protocols using nugc4
1
Establishment of Human Gastric Carcinoma Cell Lines
2
Culturing GC Cell Lines
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The GC cell lines Kato-III, NUGC4, MKN45, and MKN74 were purchased from RIKEN Cell Bank (Tsukuba, Japan) and cultured in either Roswell Park Memorial Institute 1640 medium (Sigma, St. Louis, MO) or Dulbecco’s Modified Eagle Medium (Nacalai, Japan) supplemented with 10% fetal bovine serum (Trace Scientific, Melbourne, Australia). All cells were cultured in 5% carbon dioxide at 37°C in a humidified chamber.
3
Gastric Cancer Cell Line Cultivation
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MKN7, MKN45, MKN74, HGC27, and NUGC4 human GC cell lines were purchased from the Riken Cell Bank (Tsukuba, Japan). Cells were cultured in RPMI-1640 (Nacalai Tesque, Kyoto, Japan) containing 100 μg/mL of streptomycin, 100 U/mL penicillin, and 10% FBS at 37 °C in a 5% CO2 incubator. Rabbit polyclonal anti-ANO5 antibody was obtained from Funakoshi (GTX81161) for immunohistochemical (IHC) analysis and western blotting. Mouse monoclonal anti-β-actin antibody was provided by Sigma-Aldrich (St. Louis, MO, United States) and HRP-conjugated anti-rabbit and mouse secondary antibodies by Cell Signaling Technology (Beverly, MA, United States).
4
Gastric Cancer Cell Line Characterization
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The GC cell lines MKN7, MKN45, MKN74, HGC27, and NUGC4 were purchased from the Riken Cell Bank. These cells were cultured in RPMI‐1640 (Nacalai Tesque) supplemented with 100 μg/mL of streptomycin, 100 U/mL of penicillin, and 10% fetal bovine serum (FBS). Cells were cultured at 37°C in a 5% CO2 incubator. The rabbit polyclonal anti‐ANO9 antibody used in the immunohistochemical (IHC) analysis and western blotting was purchased from Abcam (ab140087). The rabbit polyclonal anti‐PD‐L2 antibody used in the IHC analysis was purchased from ProteinTech (18251‐1‐1AP). The mouse monoclonal anti‐PD‐L1 antibody was purchased from Cell Signaling Technology (#13684), the mouse monoclonal anti‐CD8 antibody was purchased from Abcam (ab17147) and the mouse monoclonal anti‐β‐actin (ACTB) antibody was purchased from Sigma‐Aldrich. Horseradish peroxidase (HRP)‐conjugated anti‐rabbit and mouse secondary antibodies were obtained from Cell Signaling Technology. We obtained PE anti‐PD‐L1 antibody (#329706) and APC anti‐PD‐L2 antibody (#329608) from Biolegends and recombinant human PD‐1 (PE) from Abcam (ab246145). We purchased IFNα from EnoGene Biotechnology (E6L00101) and STAT3 inhibitor from Abcam (ab120952).
5
Gastric Cancer Cell Line Cultivation and Antibody Analysis
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The human GC cell lines MKN7, MKN45, MKN74, HGC27, and NUGC4 were obtained from the Riken Cell Bank (Tsukuba, Japan). These cell lines were cultured in RPMI-1640 (Nacalai Tesque, Kyoto, Japan) supplemented with 100 µg/ml of streptomycin, 100 U/ml of penicillin (Nacalai Tesque), and 10% fetal bovine serum (FBS) (Sigma-Aldrich Japan, Tokyo, Japan) in a humidified incubator at 37 °C in 5% CO 2 . The rabbit polyclonal anti-LRRC8A antibody used in the immunohistochemical (IHC) analysis was purchased from Abcam (ab-157489, Cambridge, MA, UK). The mouse monoclonal anti-LRRC8A antibody used in western blotting was purchased from Sigma-Aldrich (SAB1412855 St. Louis, MO, USA). A mouse monoclonal anti-beta-actin (ACTB) antibody was purchased from Sigma-Aldrich (A5441). Rabbit polyclonal anti-SAPK/JNK, anti-phospho-SAPK/JUN, and anti-phospho-p53 (Thr81) antibodies, a mouse monoclonal anti-p53 (1C12) antibody, and a rabbit anti-p21 (Waf1/ Cip1) antibody were purchased from Cell Signaling Technology (anti-SAPK/JNK; #9252/anti-phospho-SAPK/JNK; #9251/anti-phospho-p53 (Thr81); #2676/anti-p53 (1C12); #2524/anti-p21 (Waf1/Cip1); #2947, Beverly, MA, USA). Anti-horseradish peroxidase (HRP)-linked anti-mouse and anti-rabbit secondary antibodies were obtained from Cell Signaling Technology (anti-mouse; 7076S/anti-rabbit; 7074S).
6
Gastric Cancer Cell Line Cultivation and Characterization
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The human GC cell lines KATOIII, HGC27, NUGC4, MKN45, and MKN74 and fibroblast cell line WI-38 were obtained from the Riken Cell Bank (Tsukuba, Japan). The human normal mesothelial cell line MeT-5A (CRL-9444) was purchased from ATCC (Manassas, VA, USA). These cells were grown in RPMI-1640 medium (Nacalai Tesque, Kyoto, Japan) supplemented with 100 U/ml of penicillin, 100 µg/ml of streptomycin, and 10% fetal bovine serum (FBS). Cells were cultured in flasks or dishes in a humidified incubator at 37 °C under 5% CO 2 in air.
The monoclonal anti-NIS antibody used in the immunohistochemical analysis was obtained from Thermo Fisher Scientific (Massachusetts, USA). The rabbit monoclonal c-Jun N-terminal kinase (JNK), phosphorylated JNK, extracellular signal-regulated kinase (ERK), phosphorylated ERK, p38, phosphorylated p38, NF-κB, phospho-NF-κB, and cleaved caspase 3 antibodies were purchased from Cell Signaling Technology (Beverly, MA). Rabbit polyclonal antibodies against GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase (HRP)-conjugated anti-rabbit or mouse secondary antibodies was purchased from Cell Signaling Technology (Beverly, MA). Human Stomach Total RNA was purchased from Clontech Laboratories, Inc (Mountain View, CA).
The monoclonal anti-NIS antibody used in the immunohistochemical analysis was obtained from Thermo Fisher Scientific (Massachusetts, USA). The rabbit monoclonal c-Jun N-terminal kinase (JNK), phosphorylated JNK, extracellular signal-regulated kinase (ERK), phosphorylated ERK, p38, phosphorylated p38, NF-κB, phospho-NF-κB, and cleaved caspase 3 antibodies were purchased from Cell Signaling Technology (Beverly, MA). Rabbit polyclonal antibodies against GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase (HRP)-conjugated anti-rabbit or mouse secondary antibodies was purchased from Cell Signaling Technology (Beverly, MA). Human Stomach Total RNA was purchased from Clontech Laboratories, Inc (Mountain View, CA).
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