AKTA Explorer 100 Question: two lines from pumps A&B

[Ab-kev]

Hi,
There are two small lines leading away from Pumps A&B. Each line is hooked up one of the pumps (from what I can tell). From the literature that I've found online, I think they are called wash lines A&B. Can anybody tell me what the function of these two lines are?
Right now they are in 50mL conical tube and dispensing liquid.

Thank you,

[danko]

They wash the pump’s pistons – typically a good idea when working with salts.
Usually the inlets and the outlets are placed in the same container/bottle so that the washing liquid (often water with some ethanol) is circulated.

Best Regards

[Ab-kev]

They wash the pump’s pistons – typically a good idea when working with salts.
Usually the inlets and the outlets are placed in the same container/bottle so that the washing liquid (often water with some ethanol) is circulated.

Best Regards

Great, thank you danko.
Since I started here, there has been no liquid stored in the 50mL conical that those lines are in. Yet, they always add fluid to the conical.
Does that mean that there is a leak somewhere?
What is the risk of cross mixing buffers?

Thank you so much for your help.

[danko]

Does that mean that there is a leak somewhere?

I dont't expect that, but there is a lot of air. You'll have to prime the lines, by sucking washing liquid using a syringe, while the system's running.

No risk of buffur mixing - take my word

Best Regards

[Ab-kev]

Does that mean that there is a leak somewhere?

I dont't expect that, but there is a lot of air. You'll have to prime the lines, by sucking washing liquid using a syringe, while the system's running.

No risk of buffur mixing - take my word

Best Regards

Ok thanks danko!
Which line is the liquid coming from? Right now I am only using A-18 and two sample lines for my runs. The other lines are in:
A2 & B2 in 20% EtOH
B1 in 500mM Phosphate (never used)
A12-2MNacl
A11 in H2O w/Azide

Sorry to ask such noob questions, but I just started here and before I start changing things I want a clear understanding of what's going on. So when I do suggest things it comes from logic.

Thanks again, I really do appreciate your help danko.

[danko]

If we’re still talking about the piston washing part of the configuration, there should be 2 separate lines for that (inlet and outlet) that often are led in the same container (the liquid in that container should be replenish or better yet changed to a fresh one - once in awhile)
If you see some liquid coming out of the outlet in question, while the inlet is dty/empty, then there is some sort of mistaken setup. Follow the lines (there are many in the Explorer system – I know) and make sure there are some rationales for their dispositions.
I don’t recommend any lines that aren’t in use to e led in any sort of salt containing liquids. Here I’m thinking of the B1 line (inlet) – it shouldn’t be in the phosphate buffer if not used. Place it in 23 – 30 % ethanol/water. Better yet: Use it as a elution/gradient line (ie. 2 M NaCl). I assume (judging from the mobile phase nature) you’re working with tagged protein. The best way of using this system and most of the systems with genuine multiple pumps configuration (here 2) is to use them both and thus mix the gradient in high pressure mode.
Another thing: When starting the system (not only this particular one but generally) all the lines should be primed so that air is to be seen in any of them. Because there is a risk of introducing air in the flow from lines that aren’t in use – simply because the valve is rotating during gradient runs and the ports are passing by inlets that not necessary are in use and maybe filled with air.
As mentioned above the Explorer has a huge number of valves, ports and lines, so if I should be of more help, I’d have to be on site. Which isn’t the scope here
Do you have some support/service office nearby. Maybe they could get things in system. I wouldn’t be so sure (their service is pretty far from perfect . I know that for a fact) but that’s a path to follow. (?)
You are welcome to write back and ask questions again if something new comes up to surface.

Best Regards