![]() ![]() For this semi-dry transfer, it is very important that the gel is pre-equilibrated in transfer buffer. With increasing time, however, there is a risk of over-transfer (blew through) of the proteins through the membrane, especially for lower molecular weight (<30 kDa) proteins when using membranes with a larger pore size (0.45 µm).įor semi-dry protein transfer, the transfer sandwich is placed horizontally between two plate electrodes in a semi-dry transfer apparatus. The transfer efficiency improves with increased transfer time. Transfer efficiencies of 80–100% are achievable for proteins between 14–116 kDa. A high field option exists for a single gel, which may bring transfer time down to as little as 30 minutes, but it requires the use of high voltage (up to 200 V) or high current (up to 1.6 A) and a cooling system to dissipate the tremendous heat produced. Transfers are typically performed with an ice pack and at 4☌ to mitigate the heat produced. Multiple gels may be electrotransferred in the standard field option, which is performed either at constant current (0.1 to 1 A) or voltage (5 to 30 V) from as little as 1 hour to overnight. The supported gel sandwich is placed vertically in a tank between stainless steel/platinum wire electrodes and filled with transfer buffer. The gel is then placed in the “transfer sandwich” (filter paper-gel-membrane-filter paper), cushioned by pads and pressed together by a support grid. When performing a wet transfer, the gel is first equilibrated in transfer buffer. SureLock Tandem Midi Gel Tank: capacity for up to 2 blot modules XCell SureLock Mini-Cell: capacity for up to 2 blot modules Mini Gel Tank: capacity for up to 2 Mini Blot Modules ![]() Pre-assembled Select transfer stacks: buffer not required ![]() Electrode plates are fully separated to prevent arcing or damage.1 mini gel per blot module 1–2 blot modules per tankġ midi gel per blot module 1–2 blot modules per tank Being translucent, it allows viewing of the blot sandwich to ensure correct positioning and transfer is occurring correctly. Uniform heat dispersion across the blot sandwich ensures stable transfer times and no heat induced sample loss or transfer distortions. The electrodes, comprising platinum coated anode and stainless steel cathode, will exhibit practically no corrosion and so provide many years of trouble free use. These Semi-Dry Blotters utilise a screw down lid, which secures the blot sandwich and allows complete control of pressure ensuring even transfer. Due to very low buffer volumes typically only a few millilitres of buffer are required per transfer. Semi Dry Blotting has the added benefit of economic transfers. ![]() Each unit is compatible with their respective SynPAGE vertical gel system. All units can be used for all types of blotting: Western, Southern and Northern via uncomplicated buffer and set up procedures and are compatible with gel thicknesses from 0.25 up to 10mm without the need for additional equipment. These Semi Dry Blotters offer rapid transfer times for DNA, RNA and protein blotting typically 15 to 30 minutes. ![]()
0 Comments
Leave a Reply. |
Details
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |