Hs587t

Manufactured by American Type Culture Collection

Sourced in United States

Hs587T is a cell line derived from a breast ductal carcinoma. It is routinely used in cancer research and drug discovery studies.

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5 protocols using hs587t

Breast and Cancer Stem Cell Culturing

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MCF10A and MCF10C breast cell lines were derived at the Barbara Ann Karmanos Cancer Institute (Detroit, MI) and maintained in DMEM-F/12 medium containing 5% heat-inactivated horse serum, 10 μg/mL insulin, 20 ng/mL epidermal growth factor, 0.1 μg/mL cholera enterotoxin, and 0.5 μg/mL hydrocortisone [53 (link),54 (link)]. Breast cancer cell lines MCF7, Hs587T, and MDA231 were purchased from ATCC, and were propagated in 10% fetal bovine serum (Invitrogen, Grand Island, NY); Dulbecco’s Modification of Earle’s Media (Cellgro, Herndon, VA); 2 mM L-Glutamine (Invitrogen); 200 U Penicillin/ml; 200 μg Streptomycin/ml (Invitrogen).
Human breast cancer stem cells (BCSC: CD133+, CD44+, SSEA3/4+, Oct4+, Alkaline Phosphatase+, Aldehyde Dehydrogenase+, Telomerase+), pancreatic cancer stem cells (PCSC: CD44⁺, CD133⁺, SSEA3/4⁺, Oct4⁺, Alkaline Phosphatase⁺, Aldehyde Dehydrogenase⁺, Telomerase⁺, and Nestin⁺), and prostate cancer stem cells (PrCSC: CD44⁺, CD133⁺, SSEA3/4⁺, Oct4⁺, alkaline phosphatase⁺, aldehyde dehydrogenase⁺, and telomerase⁺) were purchased from Celprogen (San Pedro, CA), and cultured using specialized media and tissue culture plastic and matrix, to preserve their CSC phenotype, according to the manufacturer’s instructions.

Chen Z., Forman L.W., Williams R.M, & Faller D.V. (2014). Protein kinase C-delta inactivation inhibits the proliferation and survival of cancer stem cells in culture and in vivo. BMC Cancer, 14, 90.

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HER3 Neutralizing Antibody Characterization

Cited 2 times

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Anti-ErbB3 antibody RTJ2 (Abcam); anti-erbB-3/HER-3 antibody, clone 2F12 (Millipore); mouse anti-human c-erbB-3 (BD Biosciences); phospho-HER3/ErbB3 (Tyr1289) (21D3) rabbit mAb, anti-b-actin, anti-b-tubulin (Santa Cruz); anti-GAPDH, anti-AKT, anti-pAKT, anti-pERK1/2, anti-p44/42 MAP kinase (Cell Signaling Technology) were used in the study. Secondary antibodies were purchased from Jackson ImmunoResearch or Invitrogen. DNaseI-RNase free and Ribonucleoside-vanadyl complex were from NEB. Human tumor cell lines used (BT-549, BT-474, Hs587T, MCF-7, MCF10A, SKBR-3, MDA-MB-231, MDA-MB-468 and HEK293) were from ATCC and grown in RPMI or D-MEM containing 10% FBS. The HER3 neutralizing antibodes (HER3Mabs) were produced in our laboratory and described previously [20 (link), 36 (link)]. Breefly, the HER3 neutralizing antibodies HER3Mabs are a monoclonal antibodies with backbone of IgG1. HER3Mabs were expressed in HEK293 freestyle cells (Life Technologies) and purified to above 95% purity using protein A/G affinity chromatography. Antibody purity was verified by protein gel electrophoresis and antibody binding was confirmed by ELISA binding assays, flow cytometry analysis. The ability to inhibit HER3 phosphorylation upon NRG-1 activation were verified by ELISA and WBs as described by our group previously [19 (link)].

Salameh A., Fan X., Choi B.K., Zhang S., Zhang N, & An Z. (2016). HER3 and LINC00052 interplay promotes tumor growth in breast cancer. Oncotarget, 8(4), 6526-6539.

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Establishment of Murine Breast Cancer Cell Lines

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Primary murine breast cancer cell lines (P¹⁷²CC and P²⁴⁵CC) were generated by mincing breast tumors into 1-mm³ fragments, which were subjected to trypsinization for 20 min at 37°C and then cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. P53R245W (391) is an adenocarcinoma breast cancer cell originating from a germline Trp53-wm245 animal. Cells were passaged no more than 12 to 20 times before the completion of the experiments. Human breast cancer cell Hs587T, HCC38 (RRID:CVCL_1267), and HCC1395 (RRID:CVCL_1249) were purchased from American Type Culture Collection (ATCC) before performing the experiments. The other human breast tumor cells lines were purchased from MD Anderson’s Cytogenetics and Cell Authentication Core and verified by DNA fingerprinting. All cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. All cell lines were routinely confirmed by PCR to be negative for mycoplasma. CRISPR knockout clones were established by treating P¹⁷²CC or P²⁴⁵CC parental cells with AAV-mut-p53 for 48 hours and subsequently seeding one cell per well in a 96-well plate. The clones were validated with Western blotting.

Dibra D., Xiong S., Moyer S.M., El-Naggar A.K., Qi Y., Su X., Kong E.K., Korkut A, & Lozano G. (2024). Mutant p53 protects triple-negative breast adenocarcinomas from ferroptosis in vivo. Science Advances, 10(7), eadk1835.

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Breast Cancer Cell Line and Tumor Sample Collection

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BT-549 (ATCC® HTB-122™), Hs 587T (ATCC® HTB-126™), MDA-MB-436 (ATCC® HTB-130™), MDA-MB-468 (ATCC® HTB-132™), MDA-MB-231 (ATCC® HTB-26™), MDA-MB-453 (ATCC® HTB- 131™), HCC1395 (ATCC® SC-CRL-2324™), HCC38 (ATCC® CRL-2314™), MCF 10A (ATCC® CRL-10317™), AU565 (ATCC® CRL-2341™), SK-BR-3 (ATCC® HTB-30™), BT-474 (ATCC® HTB-20™), HCC1419 (ATCC® CRL-2326™), HCC1954 (ATCC® CRL-2338™), T-47D (ATCC® HTB-133™), MCF7 (ATCC® HTB-22™) and ZR-75-1 (ATCC® CRL-1500™) cells were purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA). The cells were cultured in Dulbecco’s modified Eagle medium/nutrient mixture F-12 (DMEM/ F12, Gibco, California, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco, California, USA) and 50 U/mL penicillin/ streptomycin/neomycin (Invitrogen, California, USA) in a humidified (5% CO2, 37°C) incubator. All human breast tumor samples (n = 234) were obtained as specimens from anonymous donors from Taipei Medical University Hospital, Taipei, Taiwan, as approved by the Institutional Review Board (IRB) and ethics committee of the institution (P102025). Histological inspection confirmed that all patient samples consisted of greater than 80% tumor tissue. All samples (each paired tumor tissue vs. normal tissue) were collected and categorized according to clinical characteristics, such as age.

Lee W.J., Cheng T.C., Yen Y., Fang C.L., Liao Y.C., Kuo C.C., Tu S.H., Lin L.C., Chang H.W., Chen L.C, & Ho Y.S. (2021). Tea polyphenol epigallocatechin-3-gallate inhibits cell proliferation in a patient-derived triple-negative breast cancer xenograft mouse model via inhibition of proline-dehydrogenase-induced effects. Journal of Food and Drug Analysis, 29(1), 113-127.

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Breast Cancer Cell Line Manipulation

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Human breast cancer cell lines T47D, MCF-7, Hs587T, SKBR3, BT-474, and MDA-MB-231 were obtained from the American Type Culture Collection (Manassas, VA) and authenticated using short tandem repeat analysis. The human MDA-B02 breast cancer cell line (MDA-B02) is a subpopulation of the MDA-MB-231 cell line (MDA-MB-231) that was selected for its high and selective efficiency to metastasize to bone in mice [33 (link), 51 ].
Stable silencing of ITGA5 was achieved in luciferase2-expressing MDA-MB-231 and MDA-BO2 cells (MDA-231-shITGA5 and MDA-BO2-shITGA5, respectively) by transduction with lentiviral plasmids containing hairpin shRNAs targeting ITGA5. ITGA5 was overexpressed in luciferase2-expressing MCF-7 cells (MCF-7luc2 ITGA5) using the amphotropic retroviral packaging system (Clontech).

Pantano F., Croset M., Driouch K., Bednarz-Knoll N., Iuliani M., Ribelli G., Bonnelye E., Wikman H., Geraci S., Bonin F., Simonetti S., Vincenzi B., Hong S.S., Sousa S., Pantel K., Tonini G., Santini D, & Clézardin P. (2021). Integrin alpha5 in human breast cancer is a mediator of bone metastasis and a therapeutic target for the treatment of osteolytic lesions. Oncogene, 40(7), 1284-1299.

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