R70007

Manufactured by American Type Culture Collection

The R70007 is a laboratory centrifuge designed for general-purpose applications. It features a maximum speed of 4,000 rpm and a maximum relative centrifugal force (RCF) of 2,400 x g. The centrifuge can accommodate a variety of sample sizes and tube types, making it suitable for a range of laboratory tasks. This product is intended for use in research and diagnostic settings.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) 10 cm culture dishes, by Corning (2 mentions) Hek293ft, by Thermo Fisher Scientific (2 mentions) Hygromycin b, by Thermo Fisher Scientific (1 mentions) Blasticidin s, by Thermo Fisher Scientific (1 mentions) Mouse anti flag m2 antibody, by Merck Group (1 mentions) Serum plus 2, by Merck Group (1 mentions) Hek293ft, by Caisson (1 mentions) Sprague dawley rats, by Jackson ImmunoResearch (1 mentions) Thle 2, by American Type Culture Collection (1 mentions) 5xfad mice, by Jackson ImmunoResearch (1 mentions) C57bl 6 mice, by Jackson ImmunoResearch (1 mentions) 293ft cell, by Thermo Fisher Scientific (1 mentions)

6 protocols using r70007

Culturing HEK293-FT and COS-7 Cells

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HEK293-FT (female) (ThermoFisher; R70007; RRID: CVCL_6911) and COS-7 cells (Male) (ATCC; CRL-1651; RRID: CVCL_0224) were maintained in DMEM supplemented with 10% fetal bovine serum (ThermoFisher, A3160501) and 2% penicillin/streptomycin (ThermoFisher, 15140122) at 37°C and 5.0% CO₂. Cells were passaged 2x weekly by seeding 0.5–1.0 × 10⁶ cells into 10 cm culture dishes (Corning). Cell lines were not subjected to cell authentication. Cell lines were limited to 20 passages.

Murphy J.G., Gutzmann J.J., Lin L., Hu J., Petralia R.S., Wang Y.X, & Hoffman D.A. (2022). R-type voltage-gated Ca2+ channels mediate A-type K+ current regulation of synaptic input in hippocampal dendrites. Cell reports, 38(3), 110264.

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Generation of Cas13d-expressing Cell Lines

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HEK293FT cells were acquired from Thermo Fisher Scientific (R70007) and A375 cells were acquired from ATCC (CRL-1619). HEK293FT and A375 cells were maintained at 37°C with 5% CO₂ in D10 media: DMEM with high glucose and stabilized L-glutamine (Caisson DML23) supplemented with 10% fetal bovine serum (Serum Plus II Sigma-Aldrich 14009C) and no antibiotics.
To generate doxycycline-inducible RfxCas13d-NLS HEK293FT and A375 cells, we transduced cells with a RfxCas13d-expressing lentivirus at low MOI (<0.1) and selected with 5μg/mL Blasticidin S (ThermoFisher A1113903). Single cell colonies were picked after by sparse plating. Clones were screened for Cas13d expression by western blot using mouse anti-FLAG M2 antibody (Sigma F1804).
For the GFP tiling screen RfxCas13d-expressing cells were transduced with pLentiEGFPdestabilized lentivirus at low MOI (<0.1) and selected with 100μg/ml Hygromycin B (ThermoFisher 10687010) for 2 days. Single-cell colonies were grown by sparse plating. Resistant and GFP-positive clonal cells were expanded and screened for homogenous GFP expression by flow cytometry.

Wessels H.H., Méndez-Mancilla A., Guo X., Legut M., Daniloski Z, & Sanjana N.E. (2020). Massively parallel Cas13 screens reveal principles for guide RNA design. Nature biotechnology, 38(6), 722-727.

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Developing Oral QR2 Inhibitors for Metabolic and Memory Enhancement

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In this study we created orally available QR2is with the aim to replicate genetic QR2 removal phenotypes, such as reduction of metabolic stress and memory enhancement. Inhibitor specificity for QR2 against QR1 was measured using standardized fluorescent- and orthogonal-absorption activity assays. QR2is were evaluated for in vitro activity and toxicity in QR2-expressing HEK293FT (Thermo Fisher Scientific, R70007) and THLE-2 (ATCC, CRL-2706) cells, and occlusion experiments in QR2-KO cells (Supplemental Methods). QR2i-target interaction was evaluated by crystal structure. In vivo experiments were done blind (Supplemental Methods), using randomly allocated male Sprague Dawley rats (Envigo), male C57BL/6 mice (Envigo), and male and female 5xFAD mice (The Jackson Laboratory stock no. 034840-JAX), in which PK, learning behavior, and brain pathologies were measured (see Methods Behavior, and Supplemental Methods). Group size was based on previously published data for similar experiments and the use of a power calculator (https://www.stat.ubc.ca/~rollin/stats/ssize/n2.html).

Gould N.L., Scherer G.R., Carvalho S., Shurrush K., Kayyal H., Edry E., Elkobi A., David O., Foqara M., Thakar D., Pavesi T., Sharma V., Walker M., Maitland M., Dym O., Albeck S., Peleg Y., Germain N., Babaev I., Sharir H., Lalzar M., Shklyar B., Hazut N., Khamaisy M., Lévesque M., Lajoie G., Avoli M., Amitai G., Lefker B., Subramanyam C., Shilton B., Barr H, & Rosenblum K. (2023). Specific quinone reductase 2 inhibitors reduce metabolic burden and reverse Alzheimer’s disease phenotype in mice. The Journal of Clinical Investigation, 133(19), e162120.

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Mycoplasma-free cell line cultures

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HeLa cells (ATCC #CCL-2) and 293FT cells (Invitrogen #R70007) were obtained directly from ATCC and ThermoFisher Scientific, respectively. The cells were tested by mycoplasma tests on a routine basis.

Chifor A., Choi J, & Park J. (2022). NMDA receptor-targeted enrichment of CaMKIIα improves fear memory. iScience, 25(8), 104864.

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Culturing HEK293-FT and COS-7 Cells

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Wood G.E., Dutro S.M, & Totten P.A. (1999). Target Cell Range of Haemophilus ducreyi Hemolysin and Its Involvement in Invasion of Human Epithelial Cells. Infection and Immunity, 67(8), 3740-3749.

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Cell Line Authentication & Validation

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Cell line sources: Various, e.g., HEK293FT, Thermo Fisher Cat#R70007
HEK293T-hACW2, Dr Bieniasz’s lab
Vero-E6, ATCC, Cat#CRL-1586^TMExpi293F^TM, Thermo Fisher Cat#A14527
HOS-3734, ATCC, Cat#CRL-1543^TMTZMbl, Dr. Sutton’s lab.
Cell lines were authenticated by original vendors, and re-validated in the lab as appropriate, by morphology and PCRs.
All cell lines tested negative for mycoplasma.
No commonly misidentified lines involved.

Peng L., Hu Y., Mankowski M.C., Ren P., Chen R.E., Wei J., Zhao M., Li T., Tripler T., Ye L., Chow R.D., Fang Z., Wu C., Dong M.B., Cook M., Wang G., Clark P., Nelson B., Klein D., Sutton R., Diamond M.S., Wilen C.B., Xiong Y, & Chen S. (2022). Monospecific and bispecific monoclonal SARS-CoV-2 neutralizing antibodies that maintain potency against B.1.617. Nature Communications, 13, 1638.

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