Protein ID Solutions Sample Prep Guide
33
Optional Depletion of High-Abundance Proteins with Agilent Multiple Affinity Removal Spin Cartridges 3
b Draw 4 mL of Buffer A into a 5-mL plastic Luer-Lock syringe (labeled
"A").
c Attach the syringe to the spin cartridge via the Luer-Lock adapter.
d Re-equilibrate the spin cartridge by slowly pushing Buffer A through the
resin bed.
e Leave a small aliquot of buffer on the top of the frit to prevent the resin
bed and frits from running dry.
f
The spin cartridge is now ready for the next sample.
g For storage, leave the resin bed wet with Buffer A, and leave a layer of
Buffer A above the top frit. Recap both ends of the spin cartridge tightly
and store at 4 °C.
9 Analyze the flow-through fraction(s).
a Decide whether you will analyze flow-through fractions F1 and F2
separately or combined.
·
These fractions contain the low-abundance proteins.
·
Note that F2 contains less than 10% of the total flow-through protein and
so could be omitted from the analysis.
b Prior to analysis, concentrate proteins with 5 kDa MWCO spin
concentrators (part number 5185-5991). See
Chapter 4
.
Summary :
Protein ID Solutions Sample Prep Guide 33 Optional Depletion of High-Abundance Proteins with Agilent Multiple Affinity Removal Spin Cartridges 3 b Draw 4 mL of Buffer A into a 5-mL plastic Luer-Lock syringe (labeled "A").
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