FIRST
FOLD
CREATEDBY:C.DeLaO'
THISDOCUMENTCONTAINSPROPRIETARYINFORMATION
OFAPPLIEDBIOSYSTEMSANDISTOBERETAINEDIN
CONFIDENCE.NOTTOBEREPRODUCED.
COPYRIGHTAPPLIEDBIOSYSTEMS(2000)
SPECIFICATIONSFOR:
904035REVB
INSERT,PROSORBINSTRUCTIONS
DATE1/16/01
PAGE1OF1
TOLERANCES:0.03125“
P/N904035Rev.B
01/16/01
AppliedBiosystems
©Copyright2001,AppliedBiosystems
ProSorbisatrademarkofAppleraCorporationoritssubsidiaries
intheU.S.andcertainothercountries.
ProsorbSamplePreparationCartridge
®ProductInsert
Step-By-Step Protocol:
1) If sample volume is less than 100 µL, dilute sample into 100 µL of 0.1% TFA.
2) Place ProSorb device in an appropriate test tube rack.
3) Wet the PVDF membrane in the reservoir with 10 µL of methanol.
4) Add sample to reservoir (100 to 400 µL L volume).
5) Insure that the sample reservoir has intimate contact with the absorbent and fluid transfer begins.
6) Add additional sample aliquots if necessary (change absorbent after each 750 µL).
7) Add wash solutions if necessary (change absorbent after each 750 µL).
8) Remove sample reservoir and dry PVDF membrane.
9) Punch out PVDF membrane (Figure 2).
10) For peptide samples, hold PVDF by the edge with self closing tweezers, with the convex surface up, so that the
membrane does not rest on any surfaces. Add 5 µL of dilute BioBrene+ solution and allow to dry thoroughly.
11) To the dry PVDF membrane, add 15 HL of 0.1% TFA (should not completely wet the membrane) and wick off the
surface with filter paper after 15 seconds. This will reduce the level of contaminant peaks arising from BioBrene.
12) Add 4 µL of methanol to the PVDF and allow to dry thoroughly.
Sample is now ready to be placed in the sequencer.
For Research Use Only. Not For Use In Diagnostic Procedures.
User Supplied Items:
• Methanol (HPLC grade or better)
• 0.1% Trifluoroacetic acid solution (TFA)
• Dilute BioBrene+ solution (20 µL BioBrene+ and 10 µL 0.1% TFA, added to 70 µL methanol, final conce
ntration of
BioBrene, 20 ug/µL). For use with peptides.
• PVDF membrane removal punch (P/N 401397).
Introduction:
The ProSorb™ cartridge was designed for the rapid concentration and clean-up of dilute protein and peptide samples
onto a matrix suitable for sequencing. The cartridge consists of three parts: (A), sample reservoir with attached PVDF
membrane (400 µL volume), (B) replaceable absorbent (750 µL capacity), (C) holder (Figure 1). It is recommended that
the acetonitrile concentrations of reverse-phased purified peptides be reduced (by dilution with water or 0.1% TFA) to
below 15% prior to introduction into the ProSorb cartridge. Large volume samples or washes may be accommodated as
multiple loadings into the sample reservoir (A). The absorbent needs to replaced after each 750 µL that passes through
the device. However, this may vary depending upon the sample composition. It has been found that the absorbent has a
decreased capacity for some detergent containing buffers.
5.50"
NOTES:UNLESSOTHERWISESPECIFIED
1.SOFTARTTOBESUPPLIEDBY
APPLIEDBIOSYSTEMS
2.INSERTTOBEFOLDEDASDESCRIBED
THETEXTSHOULDAPPEARONONE
SIDE.
3.AFTERTHEFIRSTFOLD.COMPANY
LOGOSHOULDAPPEARONONE
SIDE.
4.PAPERSTOCK-20#WHITECASCADE
BOND,UNLESSSPECIFIEDOTHERWISE.
FINISHEDINSERTSTOBEPACKAGEDASFOLLOWED:
1.BANDEDINGROUPSOF10
2.SHRINKWRAPPEDINPACKSOF100
3.LABELEDWITHPARTNUMBER,REVISION,P.O.#,
ANDQTY.
4.00"
NOTTOSCALE
/
