As for other colleagues, I must say that the transblot turbo system is working quite well, fast and efficient protein transfer method. The only problem is to transfer high molecular weight protein, larger than 80 kDa. We get used to stain or PVDF membrane right after the blotting and we can easily see that the efficiency is quite low compared to smaller proteins. We tested several settings and improved a bit the transfert efficiency but not enough to detect slightly expressed proteins. The price is also a problem but in term of fast, reproducibility and efficiency (for low molecular weight proteins again) is fair enough to have it.

Yes. It has worked very well for us. The consumables can be a little expensive, but the speed and quality of transfer are great. I have noticed fewer bubbles with this, but they are not always avoidable.

They should now have a bulk pack of transfer membranes, pads, and buffer that is less expensive than the single-use packs. If you plan on running a lot (and who doesn't?) This may be a better choice.

Larger proteins may not transfer quite as well, but we've had no problems with proteins up to 160 kd.

Good luck!

you can improve transfer of lerger proteins by adding SDS to the transfer buffer (0.1-0.2 %)

Me too I agree with Jason and Dhanendra. I use the Trans Blot Turbo and it works very well, also with "home-made" consumable.

:-)

We have this system in our lab and it works really well. We also have the BioRad gel imager for stain free imaging and it's a great combination. We don't tend to work on very large proteins though.

i have an experience of using Semi dry electroblotter vis-a-vis tank system.I found that Semi dry electroblotte is a better choice.

We also use this system and so far I had very poor results for proteins above 120 kDa. The bands are usually very fuzzy or smeared, does adding SDS to the transfer buffer help ? I`ve never heard this before. But all in all we are very pleased with this system also compared to other manufacturers that we use in our lab.

@Tanja, the SDS is partially stripped away from your protein during the transfer by the methanol present in the transfer buffer, this is certainly detrimental for larger proteins and thus the transfer is hampered, adding SDS can decrease this effect

I too give high marks for the Trans-Blot system. I works very well for our membrane proteins in the 15 - 100 kDa size range. We have been using them for the last 5 or 6 years. Sometimes the plastic housing cracks on the older machines but the newer models seem to have fixed that problem.

I love this machine. the only consumables we purchase are the thick pads (necessary) and pvdf (millipore). You can use standard tris-glycine transfer buffer and get transfers up to 250 kDa in 30 minutes. There are multiple formulations but I use the formulation for the Bio-Rad semi-dry apparatus and it works great. Cheers.

We got this machine around 6 months in my lab. It works great for our proteins (< 100 kDa). The only problem, as already mentioned in previous posts, is the price of the consumables...

Hi, I am also interested in this sytem. Can you buy the "thick pads" from BioRad (we have lots of PVDF) in the lab? or is there a home-made alternative?.

I would only use the Bio-Rad brand pads. I tried whatman paper but with reduced effectiveness.

Hi,

I'm also having problems with the Trans-Blot® Turbo™ Transfer System from Bio-rad using the commercial packs or the conventional materials. Some dragged spots appears even when I put just the membrane into the transfer system (without gels) only to test the transference step as you can see on the attachment. Does anyone have an ideia about what is happening?

Thank you

Hello Karine,

Are you reusing the pads?  Those spots look like a gel that has been ran too fast.

Hello Torry,

Thank you for your prompt answer!

No, I'm not reusing the pads and the post transference gels coloured by coomassie did not present that. It's something that happens on the transference step because I made a test using just membrane and pads (without gel with proteins to be transfered) and the problem persists. It seems a mechanical problem..,like a drag...

Have you any new ideia?

Greets,

Karine

The trans blot turbo does work well I must admit and I only use the biorad pads with other blotting stuff purchased separately. The one thing I am interested in though with the trans blot turbo is its durability and the price for replacements are outlandishly expensive. I was wondering if anyone else had issues with its durability such as back pins rusting and cracks with the transfer casettes?

 Hello Karine,

I am using Trans Blot Turbo from Bio-Rad to transfer the gel in western blotting, it is very nice machine.

from your attached image, it seems you loaded much amount from protein

and the filter membrane in sandwich is over wet by buffer (it should be semi-dry), after soaking by semi-dry buffer. Use the blot roller to remove air and extra buffer from the assembled layers. 

the thickness of filter membrane should be about 3 mm for each up and down sandwich layers (5 sheets from whatman paper and keep the size of filter paper and membrane the same)

before you start transfer, equilibrate the gel and membrane in semi-dry transfer buffer for 15 min. 

Hi Ahmed Saif I would lijke to know if 5 sheets from whatman paper are safe. I'm afraid to damage the trans blot system!!

As for other colleagues, I must say that the transblot turbo system is working quite well, fast and efficient protein transfer method. The only problem is to transfer high molecular weight protein, larger than 80 kDa. We get used to stain or PVDF membrane right after the blotting and we can easily see that the efficiency is quite low compared to smaller proteins. We tested several settings and improved a bit the transfert efficiency but not enough to detect slightly expressed proteins. The price is also a problem but in term of fast, reproducibility and efficiency (for low molecular weight proteins again) is fair enough to have it.

Hi, 

I want to see 180kDa protein (NR2B) and i use Trans-Blot system. LAst time, tried 25V 1.3A for 20 mins and transfer did not take place at all, then with the same gel I did 25V 1.3A for 10 mins and I still see some bands on the gel, so transfer is not compteted completely. You were mentioning about addins SDS to transfer buffer, is it just literally adding SDS to Tris-Glycine transfer buffer? I put 4 watmann paper on both sides (anote and catote) Also, although I set transfer for 25V 1.3A for 10 mins , amper reaches 0.5 maximum. how can i solve it?

Cheers!

@aysegul, yes SDS to the transfer buffer will do the job

@Viphakone, nice for you but apparently other people have problems and proteins are very heterogeneous. I agree that you don't need it every time, but for some large proteins it may help, it is known that during blotting SDS can get stripped of from the protein which results in decreased transfer

yes but slightly, 0.2-0.4%

I'll try wet transfer since last time I had no protein transfer at all. Can you suggest any transfer buffer recipe because in mine for 2tl of transfer buffer I add 400ml methanol and I think it may lead anither problems.

Bests,

@Aysegulf. Just use the running buffer provided by Biorad and add methanol to it. Works really well for me. Also hope you are using a MW ladder so you can see if no protein transfer is occurring (something is really wrong) or just large proteins are not transferring (increase current and run for a little longer). 

@dayan thanks, I'll do that:)

Hello everyone,

I have seen your answers for the trans-blot turbo. I would like to bother you a little bit (sorry). I am having problems with it maybe you can give me a clue. I am using home made tris-glicin (the one suggested by biorad), 10%gels, 50ug protein and the standard protocol in the machine. The protein ladder looks perfectly in the membrane after the transfers but when we stain the gels with comassie it seems that nothings has gone into the membrane.  Any suggestion??

thank you so much!

laura

Hello Laura,

I'm having the same issue right now. Did you find any solution? It would really help me... Thanks in advance!