Hello,
We will use Tricine SDS Page protocol to seperate our low kDa proteins. In this protocol, there are anode and cathode buffer but we can not understand how should we add these buffers ? Where should be cathode and anode buffers in tank casette ? After process, how can we reuse these buffers ?
The anode buffer should be connected to the anode, I.e positive electrode, and the cathode buffer to the cathode, I.e. negative electrode. As protein-SDS complexes are negatively charged and migrate from the top to the bottom of the gel, the cathode buffer is filled into th top tank and the anode buffer into the bottom tank of the electrophoresis device.
Hello! If you're using the Bio-RAD Tetracell system, then the cathode buffer goes inside the gasket (between the gel glasses), and the anode buffer goes outside of it. Just make sure the cathode buffer does not leak to the outside. I assemble the gasket and then leave it be for a couple of minutes to observe leakings. I've been using the protocol from Schagger (2006) and it's been working perfectly for the visualization of low molecular weight peptides.
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