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P tdtomato

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The P-tdTomato is a fluorescent protein product developed by Takara Bio. It is a red fluorescent protein derived from the Discosoma species coral. The P-tdTomato exhibits bright red fluorescence when exposed to the appropriate excitation wavelength.

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Lab products found in correlation

Puromycin, by Thermo Fisher Scientific (2 mentions) H 2db mhc class 1 sorting, by BioLegend (2 mentions) Qcxip tdtomato, by Takara Bio (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) G418 sulfate, by Harvard Bioscience (1 mentions) Lenti x bicistronic expression system, by Takara Bio (1 mentions)

3 protocols using p tdtomato

1

Generation of tdTomato and EpCAM-Expressing LL/2 Cells

Cited 2 times
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A PCR product containing the sequence of tdTomato (vector ptdTomato; #63-2531, TaKaRa Clontech) was cloned into the lentiviral expression vector pLVX-IRES-neo (LentiX-Bicistronic Expression System; #63-2181, TaKaRa Clontech) to generate a pLVX-tdTomato-IRES-Neo construct. Notably, a resistance-sequence for G418-sulfate is contained in the lentiviral expression vector. The resulting nucleotide pLVX-tdTomato-IRES-Neo was verified by Sanger sequencing and restriction enzyme digestion.12 (link) LL/2 was transfected with pLVX-tdTomato-IRES-Neo using lipofection (Lipofectamine 3000; Thermo Fisher Scientific). tdtLL/2 was enriched by cultivation in selection medium containing G418-sulfate (#A2912; Biochrom) and by repetitive FACS sorting. As previously described in detail,13 (link) tdtLL/2 cells were stably transduced with a pMXs vector containing the full-length murine EpCAM (UNIPROT entry: #Q99JW5) cDNA to generate the EpCAM-overexpressing cell line EpCAM/tdtLL/2.
Xu T., Karschnia P., Cadilha B.L., Dede S., Lorenz M., Seewaldt N., Nikolaishvili E., Müller K., Blobner J., Teske N., Herold J.J., Rejeski K., Langer S., Obeck H., Lorenzini T., Mulazzani M., Zhang W., Ishikawa-Ankerhold H., Buchholz V.R., Subklewe M., Thon N., Straube A., Tonn J.C., Kobold S, & von Baumgarten L. (2023). In vivo dynamics and anti-tumor effects of EpCAM-directed CAR T-cells against brain metastases from lung cancer. Oncoimmunology, 12(1), 2163781.
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2

Isolation and Purification of Prostate Cancer Cell Lines

Cited 1 time
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The CWR22Pc prostate cancer cell line was kindly provided by Marja Nevalainen (Dagvadorj et al., 2008 (link)). We found that this cell line contained a subpopulation of cells with fibroblast-like morphology that were human EpCAM-negative and confirmed to be of mouse origin. In order to purify tumor cells and mouse fibroblasts, we plated CWR22Pc at 400–800 cells per well (6-well) in 50% conditioned media. Numerous multi-clonal, cancer epithelial islands visually free of fibroblasts were isolated by cloning cylinders and then pooled to derive the pure epithelial subline, CWR22Pc-EP, in short 22Pc-EP. Human EpCAM-negative cancer-associated mouse fibroblasts were obtained by performing mouse specific H-2Kb and H-2Db MHC class I sorting (Biolegend #114608) and the FACs purified cancer-associated fibroblasts were termed as CWR22Pc-CAF, in short 22Pc-CAF. Purified 22Pc-EP and 22Pc-CAF cells were transduced with eGFP (SGEP-Renilla) (Fellmann et al., 2013 (link)) or tdTomato (QCXIp-tdTomato, Clontech #9136–1). tdTomato was derived from vector p-tdTomato (Clontech #632531) and cloned into the AgeI and EcoRI sites of QCXIP retroviral vector. Both were and selected with 1 μg/mL puromycin (Gibco #A1113803) for 5 days.
Zhang Z., Karthaus W.R., Lee Y.S., Gao V.R., Wu C., Russo J.W., Liu M., Mota J.M., Abida W., Linton E., Lee E., Barnes S.D., Chen H.A., Mao N., Wongvipat J., Choi D., Chen X., Zhao H., Manova-Todorova K., de Stanchina E., Taplin M.E., Balk S.P., Rathkopf D.E., Gopalan A., Carver B.S., Mu P., Jiang X., Watson P.A, & Sawyers C.L. (2020). Tumor Microenvironment-Derived NRG1 Promotes Antiandrogen Resistance in Prostate Cancer. Cancer cell, 38(2), 279-296.e9.
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3

Isolation and Purification of Prostate Cancer Cell Lines

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The CWR22Pc prostate cancer cell line was kindly provided by Marja Nevalainen (Dagvadorj et al., 2008 (link)). We found that this cell line contained a subpopulation of cells with fibroblast-like morphology that were human EpCAM-negative and confirmed to be of mouse origin. In order to purify tumor cells and mouse fibroblasts, we plated CWR22Pc at 400–800 cells per well (6-well) in 50% conditioned media. Numerous multi-clonal, cancer epithelial islands visually free of fibroblasts were isolated by cloning cylinders and then pooled to derive the pure epithelial subline, CWR22Pc-EP, in short 22Pc-EP. Human EpCAM-negative cancer-associated mouse fibroblasts were obtained by performing mouse specific H-2Kb and H-2Db MHC class I sorting (Biolegend #114608) and the FACs purified cancer-associated fibroblasts were termed as CWR22Pc-CAF, in short 22Pc-CAF. Purified 22Pc-EP and 22Pc-CAF cells were transduced with eGFP (SGEP-Renilla) (Fellmann et al., 2013 (link)) or tdTomato (QCXIp-tdTomato, Clontech #9136–1). tdTomato was derived from vector p-tdTomato (Clontech #632531) and cloned into the AgeI and EcoRI sites of QCXIP retroviral vector. Both were and selected with 1 μg/mL puromycin (Gibco #A1113803) for 5 days.
Zhang Z., Karthaus W.R., Lee Y.S., Gao V.R., Wu C., Russo J.W., Liu M., Mota J.M., Abida W., Linton E., Lee E., Barnes S.D., Chen H.A., Mao N., Wongvipat J., Choi D., Chen X., Zhao H., Manova-Todorova K., de Stanchina E., Taplin M.E., Balk S.P., Rathkopf D.E., Gopalan A., Carver B.S., Mu P., Jiang X., Watson P.A, & Sawyers C.L. (2020). Tumor Microenvironment-Derived NRG1 Promotes Antiandrogen Resistance in Prostate Cancer. Cancer cell, 38(2), 279-296.e9.
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