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Plex305

Manufactured by Thermo Fisher Scientific
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The PLEX305 is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It features a compact and modular design, enabling flexibility in configuration and easy integration into various laboratory workflows.

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Lab products found in correlation

One shot bl21 de3 chemically competent e coli, by Thermo Fisher Scientific (3 mentions) Gateway lr clonase 2 enzyme mix, by Thermo Fisher Scientific (3 mentions) Pdonr223 egfr wt, by Addgene (2 mentions) Pdonr223 egfr wt, by Thermo Fisher Scientific (2 mentions)

3 protocols using plex305

1

APEX2-tagged MAPK1 expression in HCT116 and HEK293T

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We created plasmids to express APEX2 tagged MAPK1 in HCT116 and HEK293T cells. Gateway entry clones pDONR223-MAPK1-WT (#82145), pDONAR223-PRKACA (#23495) were purchased from Addgene. pLEX-305 (#41390) lentiviral backbone vector was used and engineered to substitute the hPKG (phosphoglycerate kinase) promoter with the CMV (human cytomegalovirus) promoter plus APEX2 tag fusion with HA tags in the C-terminus of the expression gene. pDONR223_MAPK1_WT were transferred to pLEX305_C-term_APEX2 destination vector using Gateway LR Clonase II enzyme mix (Thermo Fisher) based on the provided protocol. The Gateway cloning product was transformed into One Shot BL21(DE3) chemically competent E. coli (Thermo Fisher). Ampicillin was used as the selection marker on the LB plates.
Zhang T., Fassl A., Vaites L.P., Fu S., Sicinski P., Paulo J.A, & Gygi S.P. (2022). Interrogating Kinase–Substrate Relationships with Proximity Labeling and Phosphorylation Enrichment. Journal of proteome research, 21(2), 494-506.
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2

APEX2-tagged EGFR Expression in Human Cells

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We created plasmids to express APEX2 tagged EGFR in HCT116 and HEK293T cells. Gateway entry clones pDONR223-EGFR-WT (#81926) were purchased from Addgene. pLEX-305 (#41390) lentiviral backbone vector was engineered to substitute the hPKG (phosphoglycerate kinase) promoter with the CMV (human cytomegalovirus) promoter plus APEX2 tag in the C-terminus of the expression gene. pDONR223_EGFR_WT (#81926) were transferred to pLEX305_Cterm_APEX2 destination vector using Gateway LR Clonase II enzyme mix (Thermo Fisher) based on the provided protocol. Gateway cloning product was transformed into One Shot™ BL21(DE3) chemically competent E.coli (Thermo Fisher). GFP-TFG was provided by Dr. Hideki Shibata (Nagoya University, Chikusa-ku, Nagoya, Japan).
Verdaguer M.P., Zhang T., Surve S., Paulo J.A., Wallace C., Watkins S.C., Gygi S.P, & Sorkin A. (2022). Time-resolved proximity labeling of protein networks associated with ligand-activated EGFR. Cell reports, 39(11), 110950.
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3

APEX2-tagged EGFR Expression in Human Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
We created plasmids to express APEX2 tagged EGFR in HCT116 and HEK293T cells. Gateway entry clones pDONR223-EGFR-WT (#81926) were purchased from Addgene. pLEX-305 (#41390) lentiviral backbone vector was engineered to substitute the hPKG (phosphoglycerate kinase) promoter with the CMV (human cytomegalovirus) promoter plus APEX2 tag in the C-terminus of the expression gene. pDONR223_EGFR_WT (#81926) were transferred to pLEX305_Cterm_APEX2 destination vector using Gateway LR Clonase II enzyme mix (Thermo Fisher) based on the provided protocol. Gateway cloning product was transformed into One Shot™ BL21(DE3) chemically competent E.coli (Thermo Fisher). GFP-TFG was provided by Dr. Hideki Shibata (Nagoya University, Chikusa-ku, Nagoya, Japan).
Verdaguer M.P., Zhang T., Surve S., Paulo J.A., Wallace C., Watkins S.C., Gygi S.P, & Sorkin A. (2022). Time-resolved proximity labeling of protein networks associated with ligand-activated EGFR. Cell reports, 39(11), 110950.
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