4 human NSCLC cell lines (A549, H1299, HCC827, and Calu-3) and 1 human bronchial epithelial cell line (BEAS-2B) were ordered from Procell (China) and cultured appropriately. In detail, A549 was grown in Ham's F-12K (Procell, China). H1299 and HCC827 were cultivated in RPMI-1640 (Procell, China), whereas Calu-3 was inoculated in MEM (Gibco, USA). All the media mentioned above were supplemented with 10% FBS (Procell, China) and 1% p/s (Procell, China). BEAS-2B was maintained in BEGM (Procell, China). All of them were maintained in an incubator (MG80, Kuansons, China) containing 5% CO2 at 37°C.
Hcc827
Manufactured by Procell
Sourced in China
The HCC827 is a laboratory equipment designed for temperature and humidity control. It is capable of maintaining precise temperature and humidity levels within a specified range. The core function of the HCC827 is to provide a controlled environmental chamber for various scientific and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Beas 2b, by Procell (4 mentions)
H1299, by Procell (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Nci h1299, by Procell (4 mentions)
H1975, by Procell (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Hcc827, by Thermo Fisher Scientific (3 mentions)
Nci h1299, by Thermo Fisher Scientific (3 mentions)
Calu 3, by Procell (2 mentions)
Rpmi 1640, by Procell (2 mentions)
Ham s f12k, by Procell (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Beyotime (2 mentions)
Hek293t, by Procell (2 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Procell (1 mentions)
Calu 3, by Thermo Fisher Scientific (1 mentions)
Cl 0016, by Procell (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
17 protocols using hcc827
1
Cultivation of NSCLC and Bronchial Cell Lines
2
KCTD9 Modulates LUAD Cell Response
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Human LUAD cells A549 (CL-0016) and HCC827 (CL-0094, both from Procell, Wuhan, Hubei, China) were categorized into ten groups: the control, CSE, CSE + short hairpin RNA (sh)-negative control (NC), CSE + sh-KCTD9, sh-NC, sh-KCTD9, sh-KCTD9 + dimethylsulfoxide (DMSO), sh-KCTD9 + MG132, CSE + sh-KCTD9 + sh-NC, and CSE + sh-KCTD9 + sh-TOP2A groups. A549 cells were cultured in the A549 cell-specific medium (CM-0016, Procell) containing Ham's F-12 K + 10% fetal bovine serum (FBS) + 1% penicillin/streptomycin, while HCC827 cells were cultured in the HCC827 cell-specific medium (CM-0094, Procell) containing RPMI-1640 + 10% FBS + 1% penicillin/streptomycin. Subsequently, the cells were incubated in a 5% CO2 incubator at 37 °C. Both cell lines were identified by short tandem repeat and maintained normal and healthy cell morphology. They were free of mycoplasma contamination throughout the experiment.
3
Transfection of Circular RNA in Lung Cancer Cells
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Human bronchial epithelial cells (HBE) were obtained from FUHENG Biology (Shanghai, China). PLA-801D, NCI-H1299, HCC827, NCI-H1437 and NCI-H446 cells (Procell, Wuhan, China) were maintained in RPMI-1640 (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) (Hyclone, Logan, UT, USA). A549 cells (Procell) were grown in Ham's F-12K medium (Procell) containing 10% FBS. HEK-293T cells (Zhongqiaoxinzhou, Shanghai, China) were maintained in DMEM (Gibco) containing 10% FBS. All cells were cultured in a humidified atmosphere with 5% CO2 at 37°C.
For transfection, circ_0089823 siRNAs (si-circ_0089823) and their negative control (si-NC), circ_0089823 shRNA (sh-circ_0089823) and its negative control (sh-NC) were transfected into A549 cells, while circ_0089823 over-expression plasmid (OE-circ_0089823) and its negative control (vector) were transfected into PLA-801D cells using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA) according to the protocol. Similarly, microRNA mimics, inhibitors and their negative controls (mimics NC and inhibitor NC) were transfected into A549 cell and PLA-801D cells using Lipofectamine 2000 Reagent. Cells transfected with sh-circ_0089823, sh-NC, OE-circ_0089823 or vector were selected with G418 (350–400 μg/ml; Solarbio, Beijing, China) to obtain stably transfected cell clones.
For transfection, circ_0089823 siRNAs (si-circ_0089823) and their negative control (si-NC), circ_0089823 shRNA (sh-circ_0089823) and its negative control (sh-NC) were transfected into A549 cells, while circ_0089823 over-expression plasmid (OE-circ_0089823) and its negative control (vector) were transfected into PLA-801D cells using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA) according to the protocol. Similarly, microRNA mimics, inhibitors and their negative controls (mimics NC and inhibitor NC) were transfected into A549 cell and PLA-801D cells using Lipofectamine 2000 Reagent. Cells transfected with sh-circ_0089823, sh-NC, OE-circ_0089823 or vector were selected with G418 (350–400 μg/ml; Solarbio, Beijing, China) to obtain stably transfected cell clones.
4
Isolation and Characterization of Human NSCLC Cell Lines
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Human NSCLC cell lines such as A549 (KRAS.G12S), NCI-H1299 [H1299] (NRAS Q61K), NCI-H23 [H23] (KRAS G12C), NCI-H460 [H460] (KRAS Q61H), HCC827 (EGFR del 19), and NCI-H838 [H838] (KRAS WT), human bronchial epithelial cell line HBE, and human embryonic kidney cell line HEK293T which is an excellent tool cell line for transfection and immunoprecipitation (IP) experiments were purchased from Procell (Wuhan, Hubei, China). The NSCLC cell lines and HBE cell line were cultured in Roswell Park Memorial Institute 1640 (RPMI 1640, BasalMedia, Shanghai, China). The HEK293T cell line was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, BasalMedia). The mediums were supplemented with 10% fetal bovine serum (ExCell Bio, Shanghai, China) and 1% penicillin/streptomycin (Beyotime, Nantong, Jiangsu, China).
5
NSCLC Cell Line Culture Protocol
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The cell lines used in our study, including human NSCLC cell lines (95-D, H1299, H460, HCC-827, A549, PC-9, and H1975) and human normal bronchial epithelioid cells (HBE) (Procell Life Science and Technology, Wuhan, China), were cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium containing 10% fetal bovine serum (Biological, Kibbutz Beit Haemek, Israel) with penicillin and streptomycin and incubated in an environment with 5% CO2 at 37°C.
6
Multiplex Protein Expression in Lung Cancer
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The cell lines used were as follows: HAE (CP-H209, Procell, Wuhan, China), A549 (CL-0016, Procell), HCC827 (CL-0094, Procell), NCL-H1299 (CL-0165, Procell), and NCL-H524 (CL-0403, Procell).
The following antibodies were used: anti-TMED2 (ab251705, Abcam, Cambridge, USA), anti-Ki-67 (ab15580, Abcam), anti-CEA (ab207718, Abcam), anti-NSE (ab180943, Abcam), anti-EGFR (ab200828, Abcam), anti-TLR4 (ab13556, Abcam), anti-NF-κB-p-p65 (BM3940, Boster, Wuhan, China), anti-IL-1β (ab2105, Abcam); anti-IL-18 (ab207323, Abcam); and anti-β-actin (M01263-2, Boster). The secondary antibodies used were anti-rabbit IgG (AS014, ABclonal, Wuhan, China) and anti-mouse IgG (H+L) (AS003, ABclonal).
The following antibodies were used: anti-TMED2 (ab251705, Abcam, Cambridge, USA), anti-Ki-67 (ab15580, Abcam), anti-CEA (ab207718, Abcam), anti-NSE (ab180943, Abcam), anti-EGFR (ab200828, Abcam), anti-TLR4 (ab13556, Abcam), anti-NF-κB-p-p65 (BM3940, Boster, Wuhan, China), anti-IL-1β (ab2105, Abcam); anti-IL-18 (ab207323, Abcam); and anti-β-actin (M01263-2, Boster). The secondary antibodies used were anti-rabbit IgG (AS014, ABclonal, Wuhan, China) and anti-mouse IgG (H+L) (AS003, ABclonal).
7
Cell Culture Conditions for Human Bronchial and Lung Cancer Lines
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Human bronchial epithelial cell line 16 HBE (#CL-0249) and lung cancer cell lines H1299 (#CL-0165), A549 (#CL-0016), HCC827 (#CL-0094), and H460 (#CL-0299) were obtained from Procell (Wuhan, China). Except the A549 cell line was maintained in Ham’s F-12 K medium (Thermo, Waltham, MA, USA), other cell lines were cultured in RPMI-1640 medium (Thermo). The growth conditions for these cells were in an incubator with 5% carbon dioxide at 37°C. To maintain cell growth, these media need to be supplemented with 10% FBS (Thermo) and 1% Penicillin–Streptomycin (Sigma-Aldrich, St. Louis, MO, USA).
8
Lung Adenocarcinoma Tissue and Cell Line Protocol
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This study includes five pairs of lung adenocarcinoma (LUAD) tissues from Space Central Hospital. The ethical approval for this study was from Space Central Hospital (20,200,511-JJJHZ-01). Written informed consents were obtained from each patient. Four NSCLC cell lines (A549, NCI-H1299, HCC827, and NCI-H1975) and human bronchial epithelial cell line HBEC were obtained from Procell Life Science&Technology Co.,Ltd (Wuhan, China). A549 cells were cultured in DMEM (High glucose, Gibco, Grand Island, NY, USA, 11,965–092) with 10% FBS (Gibco) and 1% penicillin/streptomycin (P/S, Procell). NCI-H1299, HCC827, and NCI-H1975 cells were cultured in RPMI-1640 medium (Gibco) with 10% FBS (Gibco) and 1% penicillin/streptomycin (P/S, Procell). HBEC cells were cultured in the bronchial epithelial growth medium (BEGM, Lonza, CC-3170). All the cells were cultured at 37 °C in a 5% CO2 incubator.
9
Cell Line Culture for Lung Cancer Research
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LUAD cell line A-549 (CRM-CCL-185) and LULC cell lines NCI-H2342 (CRL-5941) and NCI-H1299 (CRL-5803) were originally from ATCC (Manassas, VA); LUAD cell line HCC827 (CRL-2868) and human bronchial epithelioid cells 16HBE (CL-0249) were from Procell (Wuhan, China). HCC827 and NCI-H1299 cells were cultured in the commendatory RPMI-1640 medium (72400120; Gibco, Grand Island, NY), NCI-H2342 cells were in HITES medium (ATCC), and A-549 cells were in F-12K medium (21127030; Gibco, Grand Island, NY). NCI-H2342 cells were maintained in foetal bovine serum (FBS; 10100147; Gibco, Grand Island, NY) to a final concentration of 5%, and other cells were incubated in 10% FBS. All cells were in a cell culture incubator at 37 °C with 5% CO2.
10
Culturing NSCLC Cell Lines H1299 and HCC827
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Human non-small cell lung cancer (NSCLC) cell lines H1299 and HCC827 were purchased from Procell Life Science&Technology Co.,Ltd. (Wuhan, China) and cultured in Roswell Park Memorial Institute-1640 (RPMI-1640) medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA, USA). Cells were maintained at 37°C with 5% CO2.
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