Bio-Rad BioLogic DuoFlow Pathfinder 80 System DuoFlow Chromatography System Starter Kit User Manual
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2.2 Prepare Buffers
During solution preparation, wear appropriate laboratory protective clothing
including, eye protection, and gloves. Avoid skin and eye contact with starter kit
solutions. In case solutions come in contact with eyes, rinse immediately with
plenty of water and get medical advice.
Buffer A
a.
Empty the contents of the bottle labeled buffer A into a 500 ml graduated
cylinder and add filtered, high-quality water to a 500 ml volume.
b.
Place the contents of the graduated cylinder into a 1 L side-arm flask and
drop in a stirbar. Cap the side arm flask, place it on a stirplate and connect it
to a vacuum source. Degas the buffer for approximately 15 minutes with
gentle stirring.
c.
When degassing is complete, pour the buffer into a bottle and label it “Buffer A,
25 mM Tris-HCI, pH 8.1”.
Buffer B
Prepare buffer B by following the same procedure for preparation of buffer A.
Label the buffer as “Buffer B = 25 mM Tris-HCl, pH 8.1, 0.5 M NaCl”.
Conversion of Maximizer Solutions to Buffers A and B
The starter kit contains solutions for use with the DuoFlow Maximizer or
Pathfinder systems. These can be converted to buffer A, 25 mM Tris-HCI, pH 8.1,
and buffer B = 25 mM Tris-HCl, pH 8.1, 0.5 M NaCl as follows:
a. Dilute Maximizer solutions A1 and A2 to 500 ml each with filtered water.
b. Combine 150 ml of diluted A1, 100 ml of diluted A2 and the entire contents
of solution B2. Dilute the mixture to 500 ml. Check the pH and adjust to pH 8.1,
if necessary. Degas the solution and label it as “Buffer B, 25 mM Tris-HCI,
pH 8.1, 0.5 M NaCl“.
c.
Combine the remaining diluted solutions A1 and A2 with water in a 1:1:2
ratio (i.e., 250 ml each of diluted A1 and A2 with 500 ml water). Check the
pH and adjust it to pH 8.1, if necessary. Degas the solution and label it as
“Buffer A, 25 mM Tris-HCI, pH 8.1”.
2.3 Prepare Sample
a.
Remove the aluminum cap from the anion exchange standard vial. Slowly
remove the rubber plug from the vial (the contents may be under vacuum).
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