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COMPARING SDS-PAGE SYSTEMS


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7 replies to this topic

#1 self

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Posted 28 January 2011 - 02:39 PM

Hi all,

Ok, here is my first post.

We are tired of the glitches with our BioRad system (Protean III) which we have had for a while, but now combs and glass plates don't want to behave. We bought a new system from BioRad but we had immediate problems with the 'thin film of gel between glass and comb' so we returned it.

We are kind of stuck considering that there are only few selections;

1. Novex, I know from experience, is excellent. But at $10 a gel or $4 for an empty plastic cassette to pour gels is too costly. You must have guessed by now we ususally pour our own gels

2. BioRad from experience is not what it used to be....it is out for the time being.

4. Hoefer Scientific (via Fisher Sci), appears to have a very convoluted gel casting system, just looking at the manual. Has anyone had experience with Hoefer? Do gels pour well? Does the lower 'sealing door' at bottom gel really prevent leaking? AND HOW LONG HAVE YOU USED IT !!!

5. Owl Scientific. They have this pouch method for pouring gels. The pouches are $0.50 a piece, so no big deal....but pouches??? Has anyone used this system, please? Do the pouches work well? Are the puches reusable? Do gels run well? AND HOW LONG HAVE YOU USED IT !!!!!

6. There are some smaller companies...like Gbioscience and the like.

Are these the only choices? ANY IDEAS ON GEL SYSTEMS....especially IF YOU HAD LONG TERM EXPERIENCE WITH THEM is GREATLY APPRECIATED !!

Again, THANKS to any who replies....and helps our decision become less of a headache !!!

#2 bob1

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Posted 28 January 2011 - 11:26 PM

I have used Hoefer for about 6 years now and it is excellent in my opinion. It does take a bit of time to learn how to set up so that it doesn't leak, but once optimised, it has very nice gels with a good resolving distance (especially c.f. the terrible biorad system). It also has a very nice transfer unit which only takes 300 ml buffer for a wet transfer. I find that it is nice to be able to customise the thickness of the gels just using spacers and combs, rather than having to buy individual plates with built in spacers. Invitrogen pre-cast gels will fit into the gel running system too, which is handy sometimes.

I think the owl pouches will be re-useable and could probably be replaced by cheap plastic bags without buying Owl's ones. I think the pouches are only for one gel system, the ones I saw seemed to use another system, but I didn't look too closely. Owl also have aluminia plates which are good for cooling and pretty much unbreakable.

#3 Adrian K

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Posted 30 January 2011 - 02:26 AM

I use Novex, usually use for SDS-PAGE work, and for all my western blotting work.

Else, for native PAGE, I use Hoefer SE260 Mighty Small. However, I use 1.5mm spacer and comb rather than the original 1.0mm. I do have some leakage issue because I still have not learn how to set up it to avoid the leakage, yet. Used it for about 0.5-1 year.

The new bought Biorad in our lab having some leakage issue.(my colleagues use it) For me, compare with my Hoefer Mighty small, it does lose its advantages by running a relatively "shorter" gel.

Not sure about the others.

P/S: Hi Bob, any tips to avoid the leakage? I wish to learn from you... ^_^
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#4 bob1

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Posted 30 January 2011 - 04:09 PM

P/S: Hi Bob, any tips to avoid the leakage? I wish to learn from you... ^_^

Make sure that the spacers and plates align at the bottom of the casting rig - you can push the spacers down with a 200 ul tip while the screws are loose. You can confirm the alignment by looking along the bottom of the rig from the side (so that you can see both plates running like railway tracks with the spacer between them). If you have more than one set of spacers, I have found that some of the older ones are slightly less thick than some of the newer ones which means that there is a tendency to leak a little bit if they are not paired correctly.

Also make sure that the plates are square in the casting rig, so that one side isn't higher than the other if looking at it from the front.

Make sure that you clean the spacers too, we had a few the built up dried acrylamide and made it hard to cast the gels without leaks.

#5 self

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Posted 31 January 2011 - 10:02 AM

Thanks to all who replied. Input is appreciated !!

If anyone else has further input please reply.

#6 Kaioshin

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Posted 12 February 2011 - 04:37 PM

We are tired of the glitches with our BioRad system (Protean III) which we have had for a while, but now combs and glass plates don't want to behave. We bought a new system from BioRad but we had immediate problems with the 'thin film of gel between glass and comb' so we returned it.



That sounds like a problem of having too much TEMED in your stacking gel.
It's funny, the more I use it, the more I realize that problems that I've previously had and others generally have are caused by not actually knowing how to use the system properly. I'm at a point now where I never have leaks, either when casting or running a gel, nor do I have problems with polymerization and wells AND I never get any similing or frowning from my gels.

#7 proteaHeather

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Posted 16 February 2011 - 11:37 AM

Hello,
I personally only have hands on experience with the Novex and BioRad units. I have seen similar results to what is discussed above. When using BioRad precasts or hand poured glass plates, I have had issues with the running buffer leaking during the run. One solution to this problem without switching gel boxes is using a different vendor to supply the gels/cassettes. For example Protea precast gelsrequire you to reverse the gasketing to the solid side, thus eliminating the leaking issue. These precast gels are offered to be compatible with both the BioRad and Novex gel boxes and are a little cheaper per gel/cassette. My experience with the Novex unit has been good. I find that it requires a little more running buffer than the BioRad unit, but the resolving area in the gel is larger.

As far as the webbing (thin film) that you are encountering, I agree that this is due to fast polymerization rate. This can be remedied by lower the TEMED added when pouring your stacking gel. This will allow more time for the gel solution to settle from around the comb before polymerization occurs.

Hope this helps.

#8 Danio ivanio

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Posted 13 December 2011 - 04:50 AM

Hey all,

I found this topic lying there, so I wanted to ask something. We purchased the OWL System and the Casting stand with the pouches. Somebody out there who can tell me how to use it, nowhere I can find a proper description. Put spacers at bottom and on the sides and simply put it in the bag, then fix it on the stand? How is that supposed to work?

Thanks whoever replies;-)




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