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Lncap cell

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LNCaP cells are a well-characterized human prostate cancer cell line. They were originally derived from a lymph node metastasis of a human prostate adenocarcinoma. LNCaP cells express prostate-specific antigen (PSA) and are androgen-responsive.

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Lab products found in correlation

Rpmi 1640, by Thermo Fisher Scientific (19 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions) Pc3 cells, by American Type Culture Collection (11 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions) Penicillin, by Thermo Fisher Scientific (6 mentions) Streptomycin, by Thermo Fisher Scientific (6 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions) Fetal bovine serum (fbs), by Merck Group (5 mentions) Rpmi 1640 medium, by American Type Culture Collection (5 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions) Du145 cells, by American Type Culture Collection (5 mentions) Rwpe 1, by American Type Culture Collection (4 mentions) 22rv1 cell, by American Type Culture Collection (4 mentions) Vcap cells, by American Type Culture Collection (4 mentions) Rpmi 1640 medium, by Corning (4 mentions) Hek293, by American Type Culture Collection (3 mentions) Crl 1740, by American Type Culture Collection (3 mentions) Hek293t cell, by American Type Culture Collection (3 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin glutamine (psg), by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

LNCaP PCa cells (6 citations) LNCaP and PC3 cells (8 citations) LNCaP prostate cancer cells (9 citations) LNCaP and PC3 human prostate cancer cells (3 citations) LNCaP and VCaP cells (2 citations) LNCaP human prostate cancer cells (8 citations) LNCaP and 22Rv1 cells (6 citations) LNCaP prostate cancer cell line (2 citations) LNCaP and PC‐3 human prostate cancer cell lines (2 citations) PCa LNCaP and PC3 cell lines (2 citations)

130 protocols using lncap cell

1

Culturing Prostate Cancer Cell Lines

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Human prostate cancer cell line C4-2B cells were purchased from MD Anderson Characterized Cell Line Core Facility (Houston, TX), and LNCaP cells were purchased from ATCC (Manassas, VA). Both cells lines were maintained in standard cell culture conditions (5% CO2, 37 °C) and cultured in RPMI-1640 medium, 10% fetal bovine serum, and 1% antibiotic–antimycotic (Gibco).
Gdowski A.S., Lampe J.B., Lin V.J., Joshi R., Wang Y.C., Mukerjee A., Vishwanatha J.K, & Ranjan A.P. (2019). Bioinspired Nanoparticles Engineered for Enhanced Delivery to the Bone. ACS applied nano materials, 2(10), 6249-6257.
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2

Prostate Cancer Cell Culture Protocol

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LNCaP cells were purchased from ATCC and cultured in RPMI1640 supplemented with 10% FBS (Hyclone, Logan, UT or Invitrogen, Carlsbad, CA, USA). LNCaP-TGF-β1(a) cells overexpressing an HA-tagged constitutively activated TGF-β1 ligand, HA-TGF-β1(a), and control LNCaP (LNCaP-Ctrl) cells were generated as previously described [37 (link)]. Human prostate stromal cell lines HPS19I, HTS33B, and HPS33Q were prepared following a previously described protocol and cultured in Bfs medium: DMEM supplemented with 5% FBS (Hyclone or Invitrogen), 5% Nu serum (Collaborative Research, Bedford, MA), 0.5 μg/mL testosterone, 5 μg/mL insulin, 100 units/mL penicillin, and 100 μg/mL streptomycin (Sigma, St. Louis, MO, USA) [5 (link), 38 (link)].
Yang F., Chen Y., Shen T., Guo D., Dakhova O., Ittmann M.M., Creighton C.J., Zhang Y., Dang T.D, & Rowley D.R. (2014). Stromal TGF-β signaling induces AR activation in prostate cancer. Oncotarget, 5(21), 10854-10869.
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3

Validation of Prostate Cancer Assays

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LNCaP cells (ATCC) were a gift from Dr. David Jarrard and were cultured in RPMI medium (Corning) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (HyClone). LNCaP cells were harvested when confluent and frozen in aliquots in growth medium plus 10% DMSO (Fisher Scientific) at -80 ºC and quickly thawed for use in assay validation experiments. White blood cells (WBCs) for assay validation experiments were derived from healthy donor blood or from the blood of a patient with prostate cancer. WBCs were selected on CD45 positivity using magnetic LS MACS columns (Miltenyi). WBCs were frozen in aliquots in PBS plus 10% DMSO (Fisher Scientific) at -80 ºC and quickly thawed for use in assay validation experiments.
Rodems T.S., Juang D.S., Stahlfeld C.N., Gilsdorf C.S., Krueger T.E., Heninger E., Zhao S.G., Sperger J.M., Beebe D.J., Haffner M.C, & Lang J.M. (2022). SEEMLIS: a flexible semi-automated method for enrichment of methylated DNA from low-input samples. Clinical Epigenetics, 14, 37.
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4

Cell Line Maintenance for Prostate Cancer Research

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C4-2 cells were purchased from UroCorporation. PC-3 and LNCaP cells were purchased from ATCC. LNCaP-Rf, a castration-resistant subline of LNCaP was described previously [21 (link)]. LNCaP-Rf and C4-2 cells were cultured in RPMI 1640 medium supplemented with 10% charcoal-stripped fetal bovine serum (FBS) (Life Technologies) (or called androgen-depleted medium) and 100 μg/ml penicillin-streptomycin-glutamine (Life Technologies) at 37°C with 5% CO2. PC-3 and LNCaP cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) (Life Technologies).
Wang D., Ding L., Wang L., Zhao Y., Sun Z., Karnes R.J., Zhang J, & Huang H. (2015). LncRNA MALAT1 enhances oncogenic activities of EZH2 in castration-resistant prostate cancer. Oncotarget, 6(38), 41045-41055.
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5

Endothelial and Prostate Cancer Cell Culture

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HUVEC cells (Lonza, Walkersville, MD, USA) were cultured in endothelial growth medium (EGM)-plus media (Lonza, Walkersville, MD, USA) and used before passage 12. To split, the cells were detached using TryplE Express (ThermoFisher Scientific, Waltham, MA, USA), spun at 300 rpm, and resuspended in EGM-plus media. LNCaP cells were purchased from ATCC (Manassas, VA, USA) and maintained in phenol red-free RPMI 1640 medium purchased from Gibco (Gaithersburg, MD, USA) supplemented with 10% FBS and 1% Pen Strep (100 U/mL penicillin + 100 μg/mL streptomycin). Cells were cultured in 5% CO2 and 95% air at 37 °C. For hypoxia experiments, cells were placed in a hypoxia chamber (BioSpherix, Parish, NY, USA) set at 1% O2.
Harris E.M., Strope J.D., Beedie S.L., Huang P.A., Goey A.K., Cook K.M., Schofield C.J., Chau C.H., Cadelis M.M., Copp B.R., Gustafson K.R, & Figg W.D. (2018). Preclinical Evaluation of Discorhabdins in Antiangiogenic and Antitumor Models. Marine Drugs, 16(7), 241.
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6

Doxorubicin and G418 Cytotoxicity Assay

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Doxorubicin hydrochloride (Catalog No. D1515) and G418 disulfate salt (Catalog No. A1720) were obtained from Sigma. Hek293 and LNCaP cells were recently purchased form ATCC. HSG (normal human salivary gland cells) were a kind gift of Dr. Gulshan Sunavala Dossabhoy. Dulbecco’s modified Eagle’s medium (DMEM) for HEK293 cells and RPMI for LNCaP cells were purchased from Sigma Aldrich and Caisson labs. Antibodies used in this study were VDAC1 total (Catalog No. ab14734) Abcam, phospho-VDAC1 (a generous gift from Dr. Yumai Chen, UCI), Cyclin A (Catalog No. sc-271645), Cyclin B (Catalog No. sc-245), Cytochrome C (Catalog No. ab65311) Abcam, Complex COX IV (Catalog No. sc-376731), actin (Catalog No. sc-8432), GAPDH (Catalog No. 2118S) CST, Cleaved Caspase-3Asp (175) (Catalog No. 9661) CST, Alpha rabbit IgG-HRP (Catalog No. 7074S) CST. Alpha -mouse IgG-HRP (Catalog No.7076) CST.
Singh V., Khalil M.I, & De Benedetti A. (2020). The TLK1/Nek1 axis contributes to mitochondrial integrity and apoptosis prevention via phosphorylation of VDAC1. Cell Cycle, 19(3), 363-375.
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7

Culturing HeLa and LNCaP cell lines

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The human adenocarcinoma HeLa cell line was purchased from the American Type Culture Collection (Rockville, MD). Cells were maintained in Eagle’s minimum essential medium (Merck-Darmstadt, Germany, EU) with 10% of fetal bovine serum (Gibco—Thermo Fisher Scientific, USA) and 1% penicillin/streptomycin (p/s) at 37 °C with 5% CO2 humidified incubator. Human prostatic cancer LNCaP cells were also from ATCC. Cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 Medium (Gibco—Thermo Fisher Scientific, USA), supplemented with 10% fetal bovine serum (Gibco—Thermo Fisher Scientific, USA) and 1% penicillin/streptomycin (p/s) in 95% humidified air, with 5% CO2 at 37 °C. Cells were passaged once a week after trypsinization and replaced with new medium twice weekly.
Nicolosi D., Genovese C., Cutuli M.A., D’Angeli F., Pietrangelo L., Davinelli S., Petronio Petronio G, & Di Marco R. (2020). Preliminary In Vitro Studies on Corynebacterium urealyticum Pathogenetic Mechanisms, a Possible Candidate for Chronic Idiopathic Prostatitis?. Microorganisms, 8(4), 463.
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8

Cell Lines and Antiandrogens Evaluation

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Cell lines were obtained from the following: LNCaP cells from Dr. Leland Chung (Cedars-Sinai Medical Center, Los Angeles, CA, USA) in September 1993, LNCaP95 from Stephen R. Plymate (University of Washington, Seattle, WA, USA), PC3, VCaP and HEK293 were from ATCC (Manassas, VA, USA), DU145 from Victor Ling (British Columbia Cancer Agency, Integrative Oncology) in October 1998. Each cell line was maintained in RPMI-1640 (for LNCaP, LNCaP95, C4-2B), MEM (for HEK293), or DMEM (PC3, VCaP and DU145) medium supplemented with 5% or 10% FBS or charcoal-stripped serum (CSS). Enzalutamide was purchased from Omega Chem (Saint-Romuald, QC, Canada). Bicalutamide from a gift from Dr. Marc Zarenda (AstraZeneca, Cambridge, England). Ralaniten and EPI-7170 were synthesized by us.
Banuelos C.A., Ito Y., Obst J.K., Mawji N.R., Wang J., Hirayama Y., Leung J.K., Tam T., Tien A.H., Andersen R.J, & Sadar M.D. (2020). Ralaniten Sensitizes Enzalutamide-Resistant Prostate Cancer to Ionizing Radiation in Prostate Cancer Cells that Express Androgen Receptor Splice Variants. Cancers, 12(7), 1991.
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9

Cell Culture and Genetic Manipulation Protocols

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LNCaP cells were obtained from ATCC. C4-2 cells and C4-2b were obtained from MD Anderson Cancer Center, Houston, TX. LNCaP-Bic cells were a kind gift from Dr Zoran Culig (General Hospital Feldkirch). DuCaP cells were a kind gift from Philips Research (Eindhoven, The Netherlands). SKOv3 cells were a kind gift from Dr. James Brenton (University of Cambridge). LNCaP, C4-2, C4-2b and DuCaP cells were maintained at 37°C in RPMI 1640 (Invitrogen) containing L-Glutamine and supplemented with 10 % fetal bovine serum (Fbs) (Gibco) in a humidified atmosphere supplied with 5 % CO2. LNCaP-Bic cells were cultured under the same conditions but medium was supplemented with 1 μM Bicalutamide (Enzo Life Science). Cells were routinely sub-cultured 1:4 using 0.25 % Trypsin-EDTA (Invitrogen) when 80-90 % confluency was reached.
Forskolin was obtained from Sigma-Aldrich. A SIK2 inhibitor, ARN3236, was obtained from Arrien Pharmaceuticals with reported low nanomolar IC50 for SIK2 both in activity assays and cell-line experiments. [22 ] SIK2 siRNA sequences were obtained from Dharmacon. [19 (link)] The non-targeting siRNA was obtained from Dharmacon. The pCMV6-Entry-myc/flag construct was obtained from Origene. The pCMV6-Entry-WT-SIK2 -myc/flag and pCMV6-Entry-SIK2-EOS-KI-myc/flag were generated as previously described. [19 (link)]
Bon H., Wadhwa K., Schreiner A., Osborne M., Carroll T., Ramos-Montoya A., Ross-Adams H., Visser M., Hoffmann R., Ahmed A.A., Neal D.E, & Mills I.G. (2014). Salt-inducible Kinase 2 Regulates Mitotic Progression and Transcription in Prostate Cancer. Molecular cancer research : MCR, 13(4), 620-635.
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10

Mycoplasma-free LNCaP Cell Treatment

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LNCaP cells (ATCC) were confirmed to be mycoplasma free everyon month and authenticated using STR profiling. Cells were treated with 5,8,11,14-Eicosatetraynoic acid (ETYA).
Siddappa M., Wani S.A., Long M.D., Leach D.A., Mathé E.A., Bevan C.L, & Campbell M.J. (2020). Identification of transcription factor co-regulators that drive prostate cancer progression. Scientific Reports, 10, 20332.
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