During western blotting, there is no current flow (mA and Watt) during electrophoresis. i have checked the connection, ph buffer, make new buffer (1X, 2X and 5X), but still no flow of the current..can anybody help me with this kind of situation...TQ in advance.
Really need help here.
look for gas bubbles on the electrodes. If no bubbles then there is no current so either the power supply is not working...new power supply or different output connections on the one you have or maybe the electrodes are broken or corroded on the western blot apparatus. if you have a multimeter check the continuity of the leads.good luck
paul
Most likely an instrument failure or a broken cable / connector / soldering point which easily and cheaply may be fixed. If you have someone in your institution who repairs or checks equipment, have it checked there, a radio or computer technician also should be able to do it if you explain to her how the blotting apparatus works.
If not, get a simple multimeter that can measure voltage and resistance (current is an option) and check by yourself. Accuracy is not an issue in that case, any really cheap device from the night market will do.
Hi Ain,
I would check the electrodes in your chamber to make sure the copper wires are not severed or corroded and also to see if they are in close contact to the electrodes. Also, try a different power source and different plugs.
I guess you place your gel on a plate to which are connected the electrodes. This plate should be wet before placing the gel on the plate (the manufacturer may recommend a product that promotes the flow of current)
Your buffer preparation may be the issue. Are you sure you are using 1X Electrophoresis buffer?
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