Thermo Fisher Scientific
iBind Western Device
SLF1000
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I wanted to try out a system that would reduce the time and effort put into performing western blots (and I do a lot of them). I wanted to compare the performance and efficiency of the iBind system with the traditional western blotting procedures. The primary reason behind choosing this system was that it was handsfree (leaving you more time to finish other stuff) and really fast (compared to overnight primary incubation and long washes).
Western blot
Nitrocellulose membrane with proteins transferred from SDS gel
The iBind device (not the Flex) was very simple to use. It was a matter of following the protocol exactly. The card needs to be wet according to the provided diagram and the membrane needs to be placed on the card taking care to avoid bubbles with protein side facing down and low molecular weight facing the user. The wells need to be filled with the provided buffer or the antibody diluted in the buffer.
Very important to let the system sit for at least 2.5 hours. The membrane has to be placed on the card without bubbles. If you are impatient (like me) you can raise the flap on the window on the surface of the device to check the levels of buffer. the wells need to be empty before you open the casette.
The figure shows side by side comparison of the blot I did on the same membrane. I tried the iBind system first. I was impatient to see results so I let the system sit for just over an hour. The result was slightly dirtier but I guess if left to 2.5 hours the results would have been better. I then stripped the membrane and reprobed for the same GAPDH antibody at the same concentration in 5% milk using the traditional method of overnight antibody incubation 3 washes for 10 mins each in TBST followed by 1hr secondary Ab at room temperature and then 3 wases of 10mins each.
The iBind system is fast, efficient, compact, portable and gets the job done
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Really fast and handsfree system
Only one antibody can be probed at a time
I loved the system and its efficiency.
Product Name: iBind™ Western DeviceSupplier Page