BlotCycler™ FAQs: Answers to your most frequently asked questions about the BlotCycler™ Automated Western Blot Device
Have there been any tech studies to compare the accuracy and results of using the BlotCyclerTM vs. manual methods?
Multiple comparisons were performed by running two identical blots; one using BlotCyclerTM, and the other manually. In this comparison which is normally done before buying a system, the results demonstrated unequivocally that the blot quality is the same or better using BlotCyclerTM. Our instrument provides a better dynamic range of the signal generated by decreasing the background. In order to realize the full benefit of the BlotCyclerTM the existing lab protocol may need modification and optimization, especially if you are running the entire western blot hybridization at 4˚C. Precision Biosystems is happy to help you with protocol optimization.
Can I perform blocking overnight at room temperature?
While a long blocking time is good, our experience does not show any improvement in results by increasing blocking time beyond 2 hours at room temperature and 5 hours at 4˚C. However, adjusting the blocking time is the easiest way to set the end of protocol to specific time.
Can I perform a protocol for the secondary antibody only?
Yes, use the preset program that starts from the second washing step and proceeds to the second incubation and the washing steps.
Is it okay to use the pre-set program when there is no buffer in tray?
The tray should always contain buffer except for a short period of time (about 1 minute) when the solution in the tray is changing.
The video claims that a maximum of 12 blots are possible, but there are only 6 trays. How is it done?
Two blots can be placed back to back in each tray without compromising the quality of washing and sensitivity. For the configuration of 6 mini trays a maximum of 12 full size blots can be used. And, for the configuration of 4 midi trays, a maximum of 8 blots is possible.
Can a protocol be run for each blot separately at different time or do they have to run all at once?
It is possible to run two different protocols simultaneously by running one protocol for 2 or 3 blots depending on the tray size.
Please note: At this time, you cannot program each tray separately.
If I started the right side by mistake, can I stop just that side?
Yes!The BlotCyclerTM Touch (Model W5) allows you to stop either the left or the right side from the status screen. We recommend that you restart the system after interrupting the protocol.
What is minimum volume of primary antibody I can use with BlotCyclerTM?
The volume of primary antibody used depends on the size of the blot. With a smaller blot placed face-down into the trays, up to 5 ml of primary antibody can be used.
Can I run the cleaning cycle on both sides simultaneously?
To ensure thorough cleaning, you can only run the cleaning cycle on one side at a time.
What is the cleaning solution?
Cleaning solution consists of the mix of detergents at 50X concentration. Two sizes are available: CL250 (250 ml) and CL500 (500 ml).
Can I change the duration of washing?
Yes! With the BlotCyclerTM Touch, the duration of the washing cycle can be changed using the programming function. In addition, the washing time can be set from 3 minutes to 20 minutes. We recommend a washing duration of 5 minutes. Then, alter the stringency of the wash action by increasing the number of washing cycles (up to 9 times).
What is the volume of wash buffer used during washing? And, can it be changed?
The volume is fixed at about 25 ml for mini trays and 45 ml for midi trays.
Is it possible to advance to the next step with a push of a button?
No. At this time the BlotCyclerTM can only run as programmed. In order to start a different protocol you need to stop the current one and start a new one.
What type of plastic is used in the sample trays? Is it porous?
The sample trays are made from polyurethane and are not porous. Polyurethane is a unique material that offers the elasticity of rubber combined with the toughness and durability of metal. It has high resistance to strong acid and alkaline solutions and to most organic solvents used in the laboratory, including ethanol (which can be used for sterilization).
Why can’t I customize a secondary antibody-only program myself? I get an error message when I try to do so.
You cannot modify preset protocols. Please use regular programming to set up a new protocol.
I was planning to use BlotCyclerTM and found liquid in antibody containers, is something wrong?
Near the end of a normal procedure, washing buffer is pumped into the antibody container to prevent cross contamination. The liquid is removed during the cleaning cycle. If you find liquid in the container, it is likely that the cleaning cycle was not run after the previous operation. In that case, please run the cleaning cycle. It’s always good practice to run the cleaning cycle at the end of every protocol.
Is the black lid available for sale?
We have tinted lids and a light protective tank. A completely black lid is available with a custom order.
The BlotCyclerTM works well with mini sized membranes, but a high background was observed with large membranes. Are there any ways to reduce that?
There are two ways to reduce background. First, increase the number of washing cycles. Or second, increase the duration of each washing cycle. The shaking speed is high enough. Any further increase in shaking speed will not affect the washing efficiency. Also, use a blot that is sized slightly less than the tray for efficient washing (for mini gels not more than 9 cm x 7.5 cm, and for midi trays 9.5 cm x14.5 cm.
Are there any consumables to be purchased?
The only consumable required to use BlotCyclerTM is the cleaning solution!
What happens after the final washing step? Do the membranes remain submerged in the wash buffer?
Yes! To prevent the blots from drying, the wash buffer is not pumped from the tray.
If only 1 or 2 trays are used, can I conserve a wash buffer?
We provide special plugs to prevent the liquid from flowing through to the unused trays.
What do I do if there are strange messages on the display instead of the normal information (W4, BlotcyclerTM Multi only)?
First, reset the display by pressing the return key. If that does not help, you may be on a wrong screen. Press to move between the help screen and the control screen, or to move between four control screens. If you are still having a problem, restart the instrument.
How do I modify a program when the instrument has already passed the initial screen? I can only turn it on/off to see the programming screen again (W4, BlotcyclerTM Multi only)?
The easiest way address this issue is to restart the instrument.
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