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IONOVATION SCOUT 

OVERVIEW:


  Ionovation company Ionovation scout restructuring of bilayer lipid membrane electrophysiological analysis system, is one of the latest products in this field, it overcomes the traditional patch clamp series of shortcomings, whether from the technical content of products or from the perspective of the application field of the product, in the electrophysiological analysis technology has always been in a leading position, the developing direction of the electrophysiological analysis technology, is the preferred cell electrophysiological analysis laboratory experimental apparatus at home and abroad. This highly flexible desktop detection system is adapted to a variety of experimental conditions and has been used to study cell membrane ion channels, various animal and plant transporters and their organelles.

Ionovation scout technical characteristics:


√ don't need to create a gigaom block

√ the only way to measure the double seal

√ can be used to detect ion channels, vesicles and ligands on various cell membranes

√easy to carry out analysis and detection for single ion channel

√The condition can be changed on both sides of the membrane to form a double channel for analysis

             

Technology principle:


       The Ionovation Scout USES artificial lipid membranes to construct ion channels. The system is measured using two separate electrophysiological measurement Chambers. The double chamber is separated (a) by a polytetrafluoroethylene septum (a) 25 mm thick and 120 micron in diameter. The volume of the two polycarbonate cells was 1.2ml. The polycarbonate chamber has holes at the top for easy access to Ag/AgCl electrodes and injection buffer (a). When the two Chambers are filled with the appropriate buffer, the lipids dissolved in the n-decane are dispersed into the pore (b) of the teflon diaphragm, which is 120 microns in diameter. The level of the buffer is continuously lowered and elevated until the excess lipids on the teflon diaphragm are removed, and an artificial biolipid membrane (C) is formed on the teflon diaphragm with a double layer of lipid self-tissue.  

       It is then manipulated according to specific target proteins or organelles. For example, a toxin similar to the hemolysin can be added directly to the polycarbonate chamber for testing. The recombinant ion channel needs to be combined with fatty acids to form recombinant lipoproteins, and these lipoproteins are then "cut" from the biofilm to fuse with the artificial lipid membrane by ion gradient or fusion peptide method.

After the sample was prepared, the electrophysiology test was carried out. By applying different voltages to monitor the current in the cell's single ion channel, where the signal with a bandwidth of 5-10 kHz is stored on the system's hard drive, the test usually takes the form of voltage clamp mode, in which different voltages are applied to get the target signal.

Application:


1.Lipid triggers

2。Cell membrane transport

3。Membrane active agents such as synaptic nucleoprotein

4。Drug safety analysis

5.Signal transduction mode analysis

Application case


Ionovation Scout test system can be used for analysis of multiple ion channels:

1。 Measuring objects: accurate measurement of thousands of ion channels, transporters and pore activities can be achieved。

2. Software: electrophysiological data collection and analysis software Patchmaster, which USES user-friendly software to realize all measurement control; User defined experimental workflow with pre-defined protocols;

3。 Automatic operation: full-automatic equipment operation (double-layer production and verification, double-layer integrity of capacitance control, film two-side perfusion system); Two layer visualization operation;

4. Current measurement: stable low noise Ag/AgCl electrode, using salt bridge to record the current range from pA to several nA

5. Consumables: one-time bimolecular chamber can be simply and quickly replaced;

Paper list:


1. Wei? K, Neef A, Van Q, Kramer S, Gregor I, Enderlein J.Quantifying the diffusion of membrane proteins and peptides in black lipid membranes with 2-focus fluorescence correlation spectroscopy.Biophys J. 2013 Jul 16;105(2):455-62. doi: 10.1016/j.bpj.2013.06.004.

2. Weingarth M, Prokofyev A, van der Cruijsen EA, Nand D, Bonvin AM, Pongs O, Baldus M.Structural determinants of specific lipid binding to potassium channels.J Am Chem Soc. 2013 Mar 13;135(10):3983-8. doi: 10.1021/ja3119114. Epub 2013 Mar 4.

3。 Theis T, Mishra B, von der Ohe M, Loers G, Prondzynski M, Pless O, Blackshear PJ, Schachner M, Kleene R。Functional role of the interaction between polysialic acid and myristoylated alanine-rich C kinase substrate at the plasma membrane。J Biol Chem。 2013 Mar 1;288(9):6726-42。 doi: 10。1074/jbc。M112。444034。 Epub 2013 Jan 17。

4. K?stler K, Werz E, Malecki E, Montilla-Martinez M, Rosemeyer H. Nucleoterpenes of thymidine and 2'-deoxyinosine: synthons for a biomimetic lipophilization of oligonucleotides Chem Biodivers. 2013 Jan;10(1):39-61. doi: 10.1002/cbdv.201100338. 

5。 Schmidt F, Levin J, Kamp F, Kretzschmar H, Giese A, B?tzel K。 Single-channel electrophysiology reveals a distinct and uniform pore complex formed by α-synuclein oligomers in lipid membranes。 PLoS One。 2012;7(8):e42545。 doi: 10。1371/journal。pone。0042545。 Epub 2012 Aug 3。

6. Betaneli V, Petrov EP, Schwille P. The role of lipids in VDAC oligomerization. Biophys J. 2012 Feb 8;102(3):523-31. doi: 10.1016/j.bpj.2011.12.049. Epub 2012 Feb 

7。 Wei? K。, Enderlein J。 Lipid Diffusion within Black Lipid Membranes Measured with Dual-Focus Fluorescence Correlation Spectroscopy。 Chemphyschem。 2012 Mar;13(4):990-1000。

8. Werz E, Korneev S, Montilla-Martinez M, Wagner R, Hemmler R, Walter C, Eisfeld J, Gall K, Rosemeyer H. Specific DNA Duplex Formation at an Artificial Lipid Bilayer: towards a New DNA Biosensor Technology. Chem Biodivers. 2012; Feb;9(2):272-81. 

9. Schmidt F, Levin J, Kamp F, Kretzschmar H, Giese A, B?tzel K. Single-channel electrophysiology reveals a distinct and uniform pore complex formed by α-synuclein oligomers in lipid membranes. PLoS One. 2012;7(8):e42545. doi: 10.1371/journal.pone.0042545. Epub 2012 Aug 3.

10. Betaneli V, Petrov EP, Schwille P. The role of lipids in VDAC oligomerization Biophys J. 2012 Feb 8;102(3):523-31. doi: 10.1016/j.bpj.2011.12.049. Epub 2012 Feb 7.

11. Wei? K., Enderlein J. Lipid Diffusion within Black Lipid Membranes Measured with Dual-Focus Fluorescence Correlation Spectroscopy. Chemphyschem. 2012 Mar;13(4):990-1000.

12。 Werz E, Korneev S, Montilla-Martinez M, Wagner R, Hemmler R, Walter C, Eisfeld J, Gall K, Rosemeyer H。 Specific DNA Duplex Formation at an Artificial Lipid Bilayer: towards a New DNA Biosensor Technology。 Chem Biodivers。 2012; Feb;9(2):272-81

13. Erika Kovács-Bogdán, J Philipp Benz, Jürgen Soll, Bettina B?lter Tic20 forms a channel independent of Tic110 in chloroplasts BMC Plant Biol. 2011; 11: 133.

14. Honigmann A, Walter C, Erdmann F, Eggeling C, Wagner R. Characterization of horizontal lipid bilayers as a model system to study lipid phase separation. Biophys J. 2010 Jun 16;98(12):2886-94. 

15. Schneider R, Etzkorn M, Giller K, Daebel V, Eisfeld J, Zweckstetter M, Griesinger C, Becker S, Lange AThe native conformation of the human VDAC1 N terminus. Angew Chem Int Ed Engl. 2010 Mar 1;49(10):1882-5.

16. Kostka M, H?gen T, Danzer KM, Levin J, Habeck M, Wirth A, Wagner R, Glabe CG, Finger S, Heinzelmann U, Garidel P, Duan W, Ross CA, Kretzschmar H, Giese A. Single-particle characterization of iron-induced pore-forming alpha -synuclein oligomers. J Biol Chem. 2008 Feb 7. 

17. van der Laan M, Meinecke M, Dudek J, Hutu DP, Lind M, Perschil I, Guiard B, Wagner R, Pfanner N, Rehling P. Motor-free mitochondrial presequence translocase drives membrane integration of preproteins. Nat Cell Biol. 2007; 9(10):1152-9. 

18. Pagliuca C, Goetze TA, Wagner R, Thiel G, Moroni A, Parcej D. Molecular properties of Kcv, a virus encoded K+ channel. Biochemistry. 2007; 46(4):1079-90. 

19. Goetze TA, Philippar K, Ilkavets I, Soll J, Wagner R. OEP37 is a new member of the chloroplast outer membrane ion channels J Biol Chem. 2006; 281(26):17989-98. Epub 2006 Apr 19

20。 Kovermann P, Truscott KN, Guiard B, Rehling P, Sepuri NB, Muller H, Jensen RE, Wagner R, Pfanner N。 Tim22, the essential core of the mitochondrial protein insertion complex, forms a voltage-activated and signal-gated channel Mol Cell。 2002; 9(2):363-73。 

21. Meuser D, Splitt H, Wagner R, Schrempf H. Mutations stabilizing an open conformation within the external region of the permeation pathway of the potassium channel KcsA. Eur Biophys J. 2001; 30(5):385-91.

22。 Hill K, Model K, Ryan MT, Dietmeier K, Martin F, Wagner R, Pfanner N。 Tom40 forms the hydrophilic channel of the mitochondrial import pore for preproteins [see comment] Nature。 1998; 395(6701):516-21。

 

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