cell lines hek293s gnti atcc crl 3022 sf9 atcc crl  (ATCC)

Journal: Methods in enzymology

Article Title: TRPV3 Expression and Purification for Structure Determination by Cryo-EM

doi: 10.1016/bs.mie.2021.02.006

Figure Lengend Snippet:

Article Snippet: Reagent or resource Source Identifier Bacterial and Virus Strains DH10Bac Thermo Fisher Scientific 10361012 Chemicals, Peptides, and Recombinant Proteins FreeStyle ™ 293 Expression Medium Gibco 12338–018 Sf-900 ™ III Serum Free Medium Complete Gibco 12658–027 Fetal Bovine Serum Gibco 16000044 Butyric acid sodium salt Acros Organics AC263191000 Cellfectin® II Reagent Gibco 10362–100 PMSF Acros Organics 215740500 Aprotinin (from bovine lung) Sigma A1153 Leupeptin Sigma L0649 Pepstatin A Sigma P4265 Strep-Tactin® Sepharose® (strep resin) iBa 2-1201-025 Digitonin Cayman Chemical Company 14952 GDN101 Anatrace 1402423-29-3 Soybean Polar Lipid Extract Avanti Polar Lipids 541602 Bio-Beads SM-2 Bio-Rad 152–8920 Critical Commercial Assays Superose 6 10/300 GL GE Healthcare 17-5172-01 Experimental Models: Cell Lines HEK293S GnTI- ATCC CRL-3022 SF9 ATCC CRL-1711 Recombinant DNA pMSP2N2 Addgene 29520 Open in a separate window

Techniques: Recombinant, Expressing

Journal: Cell

Article Title: Voltage sensor movements during hyperpolarization in the HCN channel

doi: 10.1016/j.cell.2019.11.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, Peptides, and Recombinant Proteins DMEM/F-12 medium Gibco 11330–032 Sf-900 II SFM medium Gibco 10902–088 Freestyle 293 medium Gibco 12338–018 Lipofectamine 3000 reagent Invitrogen L3000015 Cellfectin II reagent Invitrogen 10362100 lauryl maltose neopentyl glycol Anatrace NG310 Cholesteryl hemisuccinate Anatrace CH210 Digitonin Calbiochem 300410 3’ ,5’-cyclic AMP sodium salt Sigma-Aldrich A6885 Deposited Data Human HCNI EM F186C/264C This study EMDB: EMD-20846 PDB: 6UQF Human HCN1 EM Y289D This study EMDB: EMD-20847 PDB: 6UQG Experimental Models: Cell Lines CHO-K1 ATCC CRL-9618 Sf9 ATCC CRL-1711 HEK293S GnTI − ATCC CRL-3022 Recombinant DNA Human HCN1 EM F186C/264C This study N/A Human HCN1 EM Y289D This study N/A Software and Algorithms Seriel EM Mastronarde, 2005 http://bio3d.colorado.edu/SerialEM UCSF MotionCor2 Zheng et al., 2017 https://msg.ucsf.edu/em/software/motioncor2.html Gctf Zhang, 2016 http://www.mrc-lmb.cam.ac.uk/kzhang/ RELION Scheres, 2012 ; Zivanov et al., 2019 http://www2.mrc-lmb.cam.ac.uk/relion Gautomatch Kai Zhang http://www.mrc-lmb.cam.ac.uk/kzhang/ cryoSPARC2 Punjani et al., 2017 https://cryosparc.com/ Blocres Cardone et al., 2013 https://lsbr.niams.nih.gov/bsoft/programs/blocres.html UCSF Chimera Pettersen et al., 2004 https://www.cgl.ucsf.edu/chimera COOT Emsley et al., 2010 https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot PHENIX Afonine et al., 2018 https://www.phenix-online.org MolProbity Chen et al., 2010 http://molprobity.biochem.duke.edu HELANAL-Plus Kumar and Bansal, 2012 http://nucleix.mbu.iisc.ernet.in/helanalplus/index.html PyMOL Schrodinger LLC, 2017 http://www.pymol.org Other CNBR-activated Sepharose beads GE Healthcare 17043001 Superose 6 Increase, 10/300 GL GE Healthcare 29091596 R1.2/1.3 400 mesh Au holey carbon grids Quantifoil 1210627 Open in a separate window KEY RESOURCES TABLE Structure of the HCN channel in a hyperpolarized conformation is determined S4 helix is displaced by two turns, accompanying by α to 3 10 helix transition Hyperpolarization breaks S4 into two helices, one running parallel to the membrane We propose a mechanism of electromechanical coupling in the HCN channel

Techniques: Recombinant, Software