Lately we've been running A LOT of Western blots and our trusted Life Techologies rep, who always suggests new things to try, has been pushing the new Bolt Mini Gel Tank and the Bolt gels, so we bought one of the welcome packs (gel tank, 10 gels, running buffer, sample buffer, reducing agent and standards for $390). And a week later my tech begged for another one!
I only use Life Technologies Novex gels because despite the expense of the proprietary running buffer, they are better than the Bio-Rad ones and they were the only ones which worked well for mass spec band purification. The Novex NuPAGE gels cost the same as the Bio-Rad ones, but last longer and run very straight and crisp bands.
The Bolt tank and gels are completely redesigned. It's a double-length and narrow tank where gels are run side by side, which was strange at first, but this way you can only run a gel at a time with half the buffer. The tank also has a white background behind the gel to help you better see the loading. The Bolt gels have wells cut as wedges to make loading easier and to hold more sample than the Novex gels, AND Life Technologies lowered the price from $13 to $10 for Bolt. They run way faster, so that you can be done with a 4-12% gradient run in less than an hour. And last but not least, the bands are incredibly crisp (see the pictures comparing our samples run on the Novex vs Bolt). It kind of looks like a Christmas tree...
Note on 2/6/2013: After 3 months we found only one glitch. Running the gels at 160V as recommended makes the buffer too hot and causes some random gels to start melting deforming the front. We are lowering back to 120V as for the Novex.
Remember to look at the Lab things we like page for other reviews and cool products
I only use Life Technologies Novex gels because despite the expense of the proprietary running buffer, they are better than the Bio-Rad ones and they were the only ones which worked well for mass spec band purification. The Novex NuPAGE gels cost the same as the Bio-Rad ones, but last longer and run very straight and crisp bands.
The Bolt tank and gels are completely redesigned. It's a double-length and narrow tank where gels are run side by side, which was strange at first, but this way you can only run a gel at a time with half the buffer. The tank also has a white background behind the gel to help you better see the loading. The Bolt gels have wells cut as wedges to make loading easier and to hold more sample than the Novex gels, AND Life Technologies lowered the price from $13 to $10 for Bolt. They run way faster, so that you can be done with a 4-12% gradient run in less than an hour. And last but not least, the bands are incredibly crisp (see the pictures comparing our samples run on the Novex vs Bolt). It kind of looks like a Christmas tree...
Note on 2/6/2013: After 3 months we found only one glitch. Running the gels at 160V as recommended makes the buffer too hot and causes some random gels to start melting deforming the front. We are lowering back to 120V as for the Novex.
Remember to look at the Lab things we like page for other reviews and cool products
Thanks for you comment on the Bolt gels. i am starting a new lab and will demo this soon, comparing with the surelock system that i used before. Since you have had the chance to use this system for a while now, i wonder if you can comment on the durability or quality of the materials such as the locking system and electrodes? Does the plastic seem sturdy, or is it flimsy / easy to break? Durability can only be tested over time, so i appreciate your comment !
ReplyDeleteHi, it's very sturdy and feels even more substantial than the SureLock. The electrodes are still fine after a year. The only thing is that the power leads (the little tips inserting into the power supply) are covered by little flaps that sometimes break. We never had a problem, but in my old lab where people were not careful with things they broke them a couple of times. If that happens you can rebuy them for $15 or so. You could also check with your rep whether they have fixed the problem or whether they can give you a few leads for free just in case they break.
ReplyDeleteThanks for the review. We're just starting up our lab at a new institution and are looking at the Bolt system. Our Life Tech rep gave us a free Bolt tank and wet transfer setup to try out. We are cost conscious with startup funds, so I'm wondering do you find you need to use Life Tech gel running buffers to get good results with the Bolt gels?
ReplyDeleteWe have leftover cassettes for pouring SureLock gels from our old lab - will these also fit in the Bolt system? Thanks in advance
Hi, the Bolt buffer is reusable and lasts for a long time, so in reality a $60 bottle if 20X buffer is not that expensive...especially if you think that every single gel is $10. The MES buffer is optimized for the gels, so we just leave it be. One cost cutting trick we have is actually to transfer in 10mM NaBorate, it's dirt cheap (38.1gr of NaBorate in 1L for a 10X stock) and you don't have to buy methanol.
DeleteAs for pouring gels to fit in the Bolt system, I have no idea. Maybe one day we'll have to do that, but I hate pouring gels so much that I swore I would still buy a pre-cast gel with the last $10 left in my budget. I would love to know if it worked, though...
Hello! Are you using the licor system for westerns? and if so, have you ever tried an alternative blocking buffer, not the ones they sell. How did it work for you? Thanks! Love your blog.
ReplyDeleteYes, we don't use the Licor buffer because as we were trying to optimize antibodies we were told by Cell Signaling to use 5% BSA. I have also used milk successfully in the past, but milk can give problems with clumping, slight autofluorescence, etc. BSA works well.
DeleteIn our hands, the best blocking buffer for the Li-Cor Odyssey IR system is a casein-based blocking buffer from Sigma (cat# B6429). It's sold as a 10X concentrate, and costs ~$50 for 500 ml (5L of 1X), which is actually pretty cheap for commercial blocking buffers. It works great for fluorescent and colorimetric westerns, dot blots, ELISA etc, and doesn't seem to interfere w/ anti-phospho antibodies. Also works w/ biotin-streptavidin-based detection methods. We've tried half a dozen commercial blocking buffers w/ the Li-Cor Odyssey system and it consistently gives the lowest background and highest signal-to-noise ratio. Best of all its cheap and consistent from batch to batch.
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Thanks for the tip! We do a lot of anti-phospho ab staining.
DeleteThank you for sharing your experience with Bolt gels. The data looks great and the separation between two proteins with 1 kDa difference is great.
ReplyDeleteHi. Just came across your site while trying to find the pitfalls of Bolt vs Nupage. May I ask what buffers and equipment you use for Western transfer. The Bolt transfer unit gets very hot and can dry the pvdf out. Of course, cutting back voltage was the solution. But readjusting everytime to new formats is time consuming!
ReplyDeleteAlso, what was the brief protocol for the Licor you use ?
Many thanks for your time!
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