Which is better - to make my own SDS-PAGE gels according to the available recipe, or buy the commercialised ready-made SDS-PAGE gels? Will the performance of the ready-made gels be lower than the freshly-made ones?
Deciding factors are money and time. If your lab can afford to buy ready-made, go ahead and save time. Otherwise do it yourself. Resolution is more or less same, if your hands are set with casting gels. Readymade gels comes with expiry, so plan enough gel runs before your order it.
Besides Dr. Sing mentioned, there is no significant difference in the results from self-made or ready-made gel. Once you get time, you can make several gels and preserve it at 40C in wrapped condition and when necessary, you can use these gel. By this way you can save your time.
Hi there,
The main advantage of home-made gel is that you can easily adjust the recipe for Acrylamide %. If you prefer commercial gels, you have to get a set of gels according to Acrylamide%... Or prepare the gel yourself for running a specific one...
Its true. The concern factor are the money. The result was not much different. Plus, you will develop a skill by making your own gel. I prefer to make my own. Maybe there are some small problem that might occur in the process, but by troubleshooting the gel make me understand more about the fundamental of how the gel works.
Thank you everyone! Your suggestion really helps. I do agree that by making the gel myself, I'd acquire a skill in making the gel, and I'd understand more on the physiology of the gel. I have a lot of time for this research study, so making my own gels seems to be the way to go now. Thank you.
Thank you Joel. I will keep your suggestion in mind when I decide to do proteomics with my research. I could imagine that making a gradient gel will require more efforts than the usual one.
Cost aside, I think if you can get the characteristics that fit your needs, commercial gels have advantages in practicality and reproducibility. The problems come if you find yourself needing to tinker with the gel composition to improve seperation. Purchased gels also reduce the contact with liquid acrylamide which is not nice stuff...!
One advantage of commercial gels is that you do not have to handle acrylamide, which is toxic. Use a breathing mask if you must weigh out the acrylamide powder.
Hello Stephen and Ronald. I will make sure to remember to carefully handle acrylamide. Thank you for your reminder! Now that you mentioned that, I will buy a set of breathing masks as well.
If you get experience in preparation of PAGs yourself, you can prepare your custom gels for zymography, for example.
Depends on a number of factors: cost and time probably being most important. Pouring your own gels takes more time, but the reagents are cheaper. Plus as mentioned, if you know how to make a gel, the techniques are transferable to making other types of gels. However, reproducibility tends to be better with commercial gels, particularly the Bis-Tris gels (e.g. NuPAGE novel gels) from invitrogen. The traditional Laemli system gels (e.g. Biorad ready gels) when purchased as precasts also don't last long (~ 1 month) and give worse separations than freshly poured gels. In comparison the Bis-Tris gels can last over a year in storage but require expensive MES and MOPS buffers.
Thank you Yuriy and Peter. From what I could estimate, I do not have any problems with the amount of time for my research works. So I have decided to make the SDS-PAGE gels myself.
I would agree with the opinions above about Bis-Tris gels being better for getting as precast (Sigma sell some tricine gels that have a long shelf life too). Precast has several advantages (reproducibility, quality for gradients, not having to handle acrylamide, avoid the risk of total failure, time), and it is a trade-off with time. If you are using Bis-Tris gels, the buffers for making and running the gel cost $2 per gel in any case, so there is a chemical cost to the gel.
I have always felt, all things being equal, that the critical point is cost. At some point, the value of your time in preparing the gel (etc.) exceeds the price of the pre-cast gel. Unfortunately, most manufacturers appear to be pricing their gels for big pharma, so in general it is only really economic to use pre-cast for sensitive applications like Westerns, or the nice figure for the paper.
It seems that every now and then, a manufacturer tries to break the mould, and offer pre-cast gels at a price that makes it sensible for academic labs. At present, Genscript are offering gradient gels for $4 each (http://www.genscript.com/express_plus_page_gels.html). At that price, it's cheap enough for me to get them shipped to the UK and still save massively on competitors. My lab have been really happy with these. We'll see whether the offer persists into the new year!
It depends on what you are doing, the availability of the products and the time you have to do it. The precast decrease the time of the process, and the handling of the Acrilamide, that is a very potent neurotoxin. But the home made gels are cheaper but you can introduce some manipulation errors that can affect the results.
It is better to make it yourself, it is not laborious nor time consuming and its ingredients are cheap. Just make sure the ammonium persulphate is freshly prepared and your Tris buffers for both the resolving and stacking gel are at the right pH.
I was working in a lab where we always used ready-to-use gels and we never had a problem with that. It just depends on the money the lab has. If its possible it safes a lot of time and its more healthy for you because you dont have to work with TEMED and APS.
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