SURFE²R N1
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SURFE²R N1 - 1

SURFE2R N1. Real-time measurements of electrogenic transport.

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SURFE²R N1 - 2

• Label-free electrical measurements • High signal amplification compared to • Automated recording and analysis • > 100 targets tested • ~ 100 peer reviewed papers • Transporters, pumps and channels • Native membranes or proteoliposomes • Flexible experimental design • Requires only 0.1 - 1 μg protein/sensor • Fast binding kinetics can be resolved Which targets can be measured? Fast & Easy Quality Data • Real-time data, not single point read-out Large Scope SURFE2R N1. Based on proven SSM-technology. Symporter (Sugar/H+) The SURFE2R technology allows the investigation of electrogenic...

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SURFE²R N1 - 3

A membrane preparation containing the protein of interest is adsorbed to a solid supported membrane (SSM) on a gold coated sensor chip. The protein from up to 1000 cells can be added to the 3 mm electrode, thereby generating increased signals. The protein of interest is activated by a fast solution exchange. In electrogenic transporters this generates a membrane potential, which can be measured at the electrode via capacitve coupling. Size and shape of this transient current is characterized by the type of transporter, substrate concentration and other assay conditions. The peak current is...

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SURFE²R N1 - 4

SSM-based electrophysiology: Research highlights ← Transport assay variations Different assays were performed by using concentration jumps, Δx with various compounds in background, (x) or compound gradients, (x→y). The transient currents show distinct kinetics depending on the reaction observed. Respiratory Chain Complexes → Using mitochondrial inner membrane fragments all respiratory chain complexes could be analyzed using the same sensor and different assay conditions. In addition the ATP/ADP exchanger was characterized. Watzke et al. Respiratory Chain Complexes ANT Compl. I/III/IV Compl....

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SURFE²R N1 - 5

Selection of validated targets Target Substrate Preparation & Source Plasma membrane fragments (pig kidney) Plasma membrane fragments (pig gastric mucosa) Sarcoplasmatic reticulum vesicles (rabbit muscle tissue) Synaptic vesicles (rat brain tissue) Inner mitochondrial membranes (pig heart) Plasma membrane fragments (be) Proteoliposomes (yeast expression) Resp. chain compl. H+ Inner mitochondrial membranes (pig heart) Inner mitochondrial membranes (oe in CHO cells) Purple membrane fragments (be) Synaptic vesicles (rat brain tissue) Membrane vesicles (oe in CHO cells) Membrane vesicles (oe in...

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SURFE²R N1 - 6

SURFE2R - Measure More Membrane “Using the SURFE2R N1 we have recently obtained high quality data in a very short period of time. We have developed an assay to measure the activity of ammonium transporters from the Amt protein family. There has been considerable controversy over the mechanism of ammonium transport by Amt proteins and the controversy was due to the lack of quantitative kinetic data characterizing the activity of the proteins at the single channel level. The SSM technologies allows to overcome this hurdle and we are now capable of answering very challenging functional...

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