Shp 77

Manufactured by American Type Culture Collection

Sourced in United States

The SHP-77 is a laboratory equipment product designed for cell culture applications. It functions as a rack for holding and storing cell culture flasks, dishes, and plates in a controlled environment. The core purpose of the SHP-77 is to provide a secure and organized storage solution for cell culture materials.

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Lab products found in correlation

17 protocols using shp 77

Small Cell Lung Cancer Cell Culture

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Human small cell lung cancer cells (H82, SHP77, H526, H69, and DMS273) were kindly provided by Dr. Matthew Meyerson’s Laboratory at Dana-Farber Cancer Institute, USA. H82, SHP77, H526, H69 are originally purchased from the American Type Culture Collection (ATCC) and DMS273 was originally obtained from the European Collection of Cell Cultures (ECACC).
All human small cell lung cancer cells (H82, SHP77, H526, H69, and DMS273) were cultured at 37 °C in a humid atmosphere containing 5% carbon dioxide and in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). JQ1 was purchased from Selleck Chemical (Shanghai, China), and acetylcysteine (NAC), ATRA, and RRx-001 were obtained from MedChemExpress (Shanghai, China). JQ1, ATRA, and RRx-001 were dissolved and aliquoted in DMSO (Sigma-Aldrich, Shanghai, China) and NAC was diluted in nuclease-free water.

Lv Y., Lv X., Zhang J., Cao G., Xu C., Zhang B, & Lin W. (2022). BRD4 Targets the KEAP1-Nrf2-G6PD Axis and Suppresses Redox Metabolism in Small Cell Lung Cancer. Antioxidants, 11(4), 661.

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Culturing Human Cancer Cell Lines

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Human cancer cell lines A549, H1299, H460, H69, H446, SHP-77, MCF7, and MDAMB231 were obtained from ATCC (Manassas, VA, USA). These cell lines were grown in BD T175 cell culture flasks in RPMI 1640 medium (Hyclone, UT, USA) supplemented with 10% fetal bovine serum (Gibco, USA) and antibiotics (100 IU/mL penicillin, 100 mg/mL streptomycin, and 0.25 mg/mL amphotericin; Gibco, USA) at 37°C in 5% CO2. Once the cells were 85% confluent, they were washed with PBS and subjected to trypsinization using 0.25% trypsin (Gibco, USA) to collect the cells from flasks. The cells were washed with medium and finally suspended in 50 mL of complete culture medium. Approximately 15 million cells were used to seed the lung matrix.

Mishra D.K., Miller R.A., Pence K.A, & Kim M.P. (2018). Small cell and non small cell lung cancer form metastasis on cellular 4D lung model. BMC Cancer, 18, 441.

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SCLC Tissue Sampling and Cell Culture

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MRI-guided biopsy was performed to collect both SCLC (tumor) and adjacent non-tumor lung tissues from each patient. Histopathological exams were performed to confirm all tissue samples. Human SCLC cell line SHP-77 was purchased from ATCC (USA) and cultured in 90% RPMI-1640 medium mixed with 10% FBS at 37 °C with 5% CO_2.

Luo H., Zhang Y., Qin G., Jiang B, & Miao L. (2021). LncRNA MCM3AP-AS1 sponges miR-148a to enhance cell invasion and migration in small cell lung cancer. BMC Cancer, 21, 820.

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SCLC Cell Line Characterization and YAP1 Inhibition

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The SCLC cell lines SBC-3, H446, H69, H82 and SHP77 were purchased from ATCC, and DMS273 was purchased from AcceGen Biotech. Six human SCLC cell lines were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum at 37°C in a 5% CO2 incubator.20 (link) Selleck Chemicals (Shanghai, China) provided the YAP1 inhibitor (CA3). Antibodies against YAP1 and GAPDH were obtained from Abways Technology (Shanghai, China) and antibodies against CD63 and TSG101 were obtained from Cell Signaling Technology (United States).

Chao F., Zhang Y., Lv L., Wei Y., Dou X., Chang N., Yi Q, & Li M. (2023). Extracellular Vesicles Derived circSH3PXD2A Inhibits Chemoresistance of Small Cell Lung Cancer by miR-375-3p/YAP1. International Journal of Nanomedicine, 18, 2989-3006.

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Screening Anti-DLL3 Monoclonal Antibodies

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Example 1

The monoclonal antibodies to be used in accordance with the invention may be made by the hybridoma method first described by Kohler and Milstein, Nature 256:495, 1975, or may be made by recombinant DNA methods such as described in U.S. Pat. No. 4,816,567. Anti-DLL3 antibodies were first screened in Flag-DLL3 (adipogen) ELISA and then screened in FACS to determine binding to HEK-293T cells with or without human DLL3 expression.

To test if the DLL3 specific antibodies can recognize cells that express endogenous DLL3, DMS 273 (Sigma, cat #95062830), DMS 454 (Sigma, cat #95062832), and SHP-77 (ATCC, cat # CRL-2195) cells were stained with 2 ug/ml of purified DLL3 antibodies with mouse IgG2A backbone (mIgG2a) or control mIgG2a antibody in PBS supplemented with 1% BSA. Bound DLL3 antibodies were detected with PE labelled anti-mouse IgG antibody (Biolegend, cat #405307). The samples were analyzed by flow cytometry. Representative images showing binding of DLL3 antibodies to DMS 273, DMS 454 and SHP-77 cells are included in FIG. 1.

US11673953B2. DLL3 targeting chimeric antigen receptors and binding agents (2023-06-13). ALLOGENE THERAPEUTICS, INC. [US], PFIZER INC. [US]. Inventors: Yi Zhang [US], Thomas John Van Blarcom [US], Siler Panowski [US], Silvia K. Tacheva-Grigorova [US], Barbra Johnson Sasu [US].

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Lurbinectedin Treatment on Lung Cancer Cell Lines

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The following cell lines were obtained from the ATCC: A549 (lung adenocarcinoma; CCL‐185), DMS‐53 (small‐cell lung carcinoma; CRL‐2062), IMR‐90 (normal lung; CCL‐186), NCI‐H69 (small‐cell lung carcinoma; HTB‐119), NCI‐H82 (small‐cell lung carcinoma; HTB‐175), NCI‐H146 (small‐cell lung carcinoma; HTB‐173), NCI‐H460 (large cell lung carcinoma; HTB‐177), NCI‐H510A (small‐cell lung carcinoma; HTB‐184), NCI‐H526 (small‐cell lung carcinoma; CRL‐5811), and SHP‐77 (small‐cell lung carcinoma; CRL‐2195). The cells were authenticated and tested for mycoplasma contamination. All cell lines were cultured in the medium and conditions recommended by the supplier and supplemented with 10%FBS, 2 nmol/L l‐glutamine, and penicillin–streptomycin mix (Sigma). For lurbinectedin treatment, cells were seeded and grown to subconfluency before the addition of the drug to the culture medium after having optimized drug concentration (50 nM) and time of lurbinectedin treatment (4 h).

Costanzo F., Martínez Diez M., Santamaría Nuñez G., Díaz‐Hernandéz J.I., Genes Robles C.M., Díez Pérez J., Compe E., Ricci R., Li T., Coin F., Martínez Leal J.F., Garrido‐Martin E.M, & Egly J.M. (2022). Promoters of ASCL1‐ and NEUROD1‐dependent genes are specific targets of lurbinectedin in SCLC cells. EMBO Molecular Medicine, 14(4), e14841.

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Mycoplasma-free Cell Line Validation

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Chan J.M., Quintanal-Villalonga Á., Gao V.R., Xie Y., Allaj V., Chaudhary O., Masilionis I., Egger J., Chow A., Walle T., Mattar M., Yarlagadda D.V., Wang J.L., Uddin F., Offin M., Ciampricotti M., Qeriqi B., Bahr A., de Stanchina E., Bhanot U.K., Lai W.V., Bott M.J., Jones D.R., Ruiz A., Baine M.K., Li Y., Rekhtman N., Poirier J.T., Nawy T., Sen T., Mazutis L., Hollmann T., Pe’er D, & Rudin C.M. (2021). Signatures of plasticity, metastasis, and immunosuppression in an atlas of human small cell lung cancer. Cancer cell, 39(11), 1479-1496.e18.

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PLCG2 Manipulation in SCLC Cells

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H82 (male), SHP-77 (male), H526 (male), H446 (male) and DMS-114 (male) were purchased from ATCC, authenticated through the STR characterization method and regularly tested for Mycoplasma. Both cell lines were cultured in RPMI 1640 supplemented with 10% FBS and cultured according to ATCC guidelines.
Lentiviral plasmids were used for PLCG2 overexpression (GeneCopoeia, #EX-A8643-Lv201) and for PLCG2 CRISPR knock out (Sigma-Aldrich, #HSPD0000031727). Lentiviral particles were produced by standard protocols, transfecting HEK293T cells using JetPrime reagent (Polyplus, #114–15) and concentrated viruses using Lenti-X Concentrator (Takara Bio, #631232) and SCLC cells were transduced at high multiplicity of infection in a spin transduction protocol (Centrifugation of cells at 800×g, 30 minutes with 8ug/mL polybrene).

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NSCLC and SCLC Cell Lines Treatment

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Four NSCLC cell lines (H1650, PC9, H1299, and A549) and nine SCLC cell lines (DMS79, H2171, H1694, SHP77, H82, H446, H69, SW1271, and H841) were purchased from ATCC (Manassas, VA, USA) and cultured in DEME/F12 or RPMI‐1640 with 10% FBS and 5% GlutaMAX. Cells were revalidated by STR profiling if used for more than 3 years.
Trichostatin A (TSA) (Cell Signaling Technology, Danvers, MA, USA, #9950) was prepared in DMSO as a stock solution of 4 mm. Cell lines H1299 and SW1271 were seeded in 100 mm plates and treated by TSA at 400 nm and 2 mm, respectively. Cells were harvested after six hours of treatment and lysed for western blotting.

He T., Wildey G., McColl K., Savadelis A., Spainhower K., McColl C., Kresak A., Tan A.C., Yang M., Abbas A, & Dowlati A. (2020). Identification of RUNX1T1 as a potential epigenetic modifier in small‐cell lung cancer. Molecular Oncology, 15(1), 195-209.

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Cell Line Authentication and Mycoplasma Testing

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H69 (HTB-119), H82 (HTB-175), SHP-77 (CRL-2195), H526 (CRL-5811), H446 (HTB-171) and DMS-114 (CRL-2066) were purchased from ATCC, authenticated through the STR characterization method and regularly tested for mycoplasma (Universal Mycoplasma Detection Kit, #30-1012K, ATCC, last date: June 2021). Where possible all experiments were performed in low passage cells. All cell lines were cultured in RPMI 1640 supplemented with 10% FBS and cultured according to ATCC guidelines.

Quintanal-Villalonga A., Taniguchi H., Hao Y., Chow A., Zhan Y.A., Chavan S.S., Uddin F., Allaj V., Manoj P., Shah N.S., Chan J.M., Offin M., Ciampricotti M., Ray-Kirton J., Egger J., Bhanot U., Linkov I., Asher M., Roehrl M.H., Qiu J., de Stanchina E., Hollmann T.J., Koche R.P., Sen T., Poirier J.T, & Rudin C.M. (2021). Inhibition of XPO1 sensitizes small cell lung cancer to first- and second-line chemotherapy. Cancer research, 82(3), 472-483.

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