T7 coupled reticulocyte lysate system

Manufactured by Promega

The T7-coupled reticulocyte lysate system is a cell-free in vitro translation system that uses T7 RNA polymerase to drive the expression of recombinant proteins from T7 promoter-driven DNA templates. It provides a convenient platform for the rapid synthesis of proteins for various applications.

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Lab products found in correlation

Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions) Hygromycin, by Thermo Fisher Scientific (2 mentions) Tetracycline, by Merck Group (2 mentions) Chemiluminescent hrp substrate, by Merck Group (2 mentions) Glutathione sepharose bead, by GE Healthcare (2 mentions) Ponceau s, by Merck Group (2 mentions) Mg132, by Merck Group (2 mentions) Metafectene pro, by Biontex (2 mentions) Complete edta free protease inhibitor, by Roche (2 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions) Bafa1, by Santa Cruz Biotechnology (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Doxycycline, by Merck Group (1 mentions) Gfp trap, by Proteintech (1 mentions) Pen strep, by Merck Group (1 mentions) Hanks balanced salt solution (hbss), by Merck Group (1 mentions) Bafilomycin a1, by Santa Cruz Biotechnology (1 mentions) 35s methionine, by PerkinElmer (1 mentions)

2 protocols using t7 coupled reticulocyte lysate system

In Vitro Protein Synthesis and Inhibition

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Reagents and chemicals used were BafA1 (sc-201550; Santa Cruz Biotech), MG132 (C2211; Sigma-Aldrich), [³⁵S]methionine (NEG709A500UC; PerkinElmer), T7-coupled reticulocyte lysate system (14610; Promega), Ponceau S (P3504; Sigma-Aldrich), DMEM (D6046; Sigma-Aldrich), bicarbonate buffered HBSS (H9269; Sigma-Aldrich), Hygromycin (10687-010; Thermo Fisher Scientific), tetracycline (87128; Sigma-Aldrich), penicillin/streptomycin (P4333; Sigma-Aldrich), Metafectene Pro (T040; Biontex), (S0615; Biochrom), Lipofectamine RNAiMAX (13778; Thermo Fisher Scientific), cOmplete EDTA-free Protease inhibitor (11836170001; Roche), chemiluminescent HRP substrate (Sigma-Aldrich), and glutathione sepharose beads (17-5132-01; GE Healthcare).

Nthiga T.M., Shrestha B.K., Bruun J.A., Larsen K.B., Lamark T, & Johansen T. (2021). Regulation of Golgi turnover by CALCOCO1-mediated selective autophagy. The Journal of Cell Biology, 220(6), e202006128.

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Investigating VAPA and VAPB in Protein Folding

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Bafilomycin A1 (Santa Cruz Biotech sc‐201550), MG132 (Sigma‐Aldrich #C2211), [35S] methionine (PerkinElmer NEG709A500UC), T7‐coupled reticulocyte lysate system (Promega #14610), Ponceau S (sigma #P3504), Dulbeco's modified Eagle's medium (DMEM) (Sigma‐Aldrich #D6046), HBSS (Sigma‐Aldrich #H9269), Hygromycin (Thermo Fisher #10687‐010), Tetracycline (Sigma‐Aldrich #87128), Doxycycline (Sigma‐Aldrich #D9891), Pen/Strep (Sigma‐Aldrich #P4333), Metafectene Pro (Biontex #T040), fetal bovine serum (FBS) (Biochrom #S0615), Lipofectamine RNAiMAX (Thermo Fisher #13778), Complete EDTA‐free Protease Inhibitor (Roche #11836170001), Chemiluminescent HRP substrate (Sigma‐Aldrich), GFP‐TRAP (Chromotek #GA‐20), Glutathione Sepharose beads (GE Healthcare #17‐5132‐01). The following siRNA oligonucleotides were used: CALCOCO1; 5′‐GAAGCUGAGUGCAGAGAUA‐3′ (Sigma), VAPA; 5′‐CCUGAGAGAUGAAGGUUUA‐3′ (Sigma), VAPB; 5′‐GGAAGACAGUGCAGAGCAA‐3′ (Sigma), Smartpool siGENOME VAPA siRNA (Dharmacon, M‐021382‐01‐0005) and Smartpool siGENOME VAPB siRNA (Dharmacon, M‐017795‐00‐0005).

Nthiga T.M., Kumar Shrestha B., Sjøttem E., Bruun J., Bowitz Larsen K., Bhujabal Z., Lamark T, & Johansen T. (2020). CALCOCO1 acts with VAMP‐associated proteins to mediate ER‐phagy. The EMBO Journal, 39(15), e103649.

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