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Hek293t cell line crl 11268

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The HEK293T cell line (CRL-11268) is a well-established and widely used human cell line derived from human embryonic kidney cells. This cell line is known for its high transfection efficiency and is commonly used in a variety of applications, including protein expression, virus production, and gene expression studies.

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Lab products found in correlation

Pfastbac, by Thermo Fisher Scientific (2 mentions) Jm109 de3 cells, by Promega (2 mentions) Rosetta 2 de3 cells, by Merck Group (2 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions)

Close spelling variants of this product

HEK293T (CRL-11268), CRL-11268 HEK293T cell line HEK293T line HEK293T cells HEK293T

3 protocols using hek293t cell line crl 11268

1

Heterologous Expression and Purification of Ric-8A

Cited 1 time
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Rat GST-TEV-Ric-8A was expressed in Tni (Trichoplusia ni) cells infected with recombinant baculovirus (pFastBac, Invitrogen). Tni cells were obtained from Expression Systems, LLC. Rat GST-TEV-Ric-8A together with human Gαq were expressed in Tni (Trichuplusia ni) cells infected with two recombinant baculoviruses (pFastBac, Invitrogen). Human 6His-3C-Gαi1 WT or 6His-3C-Gαi1 ΔF354 were expressed in Rosetta 2(DE3) cells (Merck Millipore). Human Gαi1 (6His between M119 and T120) was expressed in JM109 (DE3) cells (Promega). MBP-Ric-8A (423–530) was expressed in Rosetta 2 (DE3) cells. The HEK293T cell line (CRL-11268) was obtained from the American Type Culture Collection (ATCC). Crispr/Cas9 procedures were used previously to delete RIC-8A in the HEK293T cells (Papasergi-Scott et al., 2018 (link)). 3X-FLAG-Ric-8A point mutant plasmids were transfected and selected for stable expression with Hygromycin B.
Seven A.B., Hilger D., Papasergi-Scott M.M., Zhang L., Qu Q., Kobilka B.K., Tall G.G, & Skiniotis G. (2020). Structures of Gα Proteins in Complex with Their Chaperone Reveal Quality Control Mechanisms. Cell reports, 30(11), 3699-3709.e6.
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2

Heterologous Expression and Purification of Ric-8A

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Rat GST-TEV-Ric-8A was expressed in Tni (Trichoplusia ni) cells infected with recombinant baculovirus (pFastBac, Invitrogen). Tni cells were obtained from Expression Systems, LLC. Rat GST-TEV-Ric-8A together with human Gαq were expressed in Tni (Trichuplusia ni) cells infected with two recombinant baculoviruses (pFastBac, Invitrogen). Human 6His-3C-Gαi1 WT or 6His-3C-Gαi1 ΔF354 were expressed in Rosetta 2(DE3) cells (Merck Millipore). Human Gαi1 (6His between M119 and T120) was expressed in JM109 (DE3) cells (Promega). MBP-Ric-8A (423–530) was expressed in Rosetta 2 (DE3) cells. The HEK293T cell line (CRL-11268) was obtained from the American Type Culture Collection (ATCC). Crispr/Cas9 procedures were used previously to delete RIC-8A in the HEK293T cells (Papasergi-Scott et al., 2018 (link)). 3X-FLAG-Ric-8A point mutant plasmids were transfected and selected for stable expression with Hygromycin B.
Seven A.B., Hilger D., Papasergi-Scott M.M., Zhang L., Qu Q., Kobilka B.K., Tall G.G, & Skiniotis G. (2020). Structures of Gα Proteins in Complex with Their Chaperone Reveal Quality Control Mechanisms. Cell reports, 30(11), 3699-3709.e6.
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3

HEK293T Cell Culture and EV Isolation

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HEK293T cell line (CRL-11268) was purchased from American Type Culture Collection (Manassas, VA). HEK293T cells were cultured as adherent cells in T-flasks or 150 mm dishes using Dulbecco’s Modified Eagle Medium (Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS) (Sigma, St. Louis, MO). The collection of EVs from cells cultured used FBS that was first depleted of exogenous EVs by an 18 h centrifugation at 100,000 rcf, 4°C. Cells were cultured in a 5% CO2/humidified air incubator. HEK293T cells were passaged using trypsin/EDTA and only passages less than 35 were used throughout the study. Cultures were scaled up to 12 or 24, 150 mm dishes prior to isolating the EVs.
Sutaria D.S., Jiang J., Elgamal O.A., Pomeroy S.M., Badawi M., Zhu X., Pavlovicz R., Azevedo-Pouly A.C., Chalmers J., Li C., Phelps M.A, & Schmittgen T.D. (2017). Low active loading of cargo into engineered extracellular vesicles results in inefficient miRNA mimic delivery. Journal of Extracellular Vesicles, 6(1), 1333882.
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