LC Columns and Accessories >> Bio Columns Selection Guide

121 Pages·2012·11.38 MB·English

Columns for proteomics Using highly pure chromatographic media and IntegraFrit columns have an integral high L
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biocompatible, metal-free fused silica capillaries, porosity frit, behind which is the packed
Nano, capillary and micro columns
EASY-Column™ capillary LC columns are produced chromatography bed. The frit end of the fused C
Acclaim PepMap and Acclaim PepMap RSLC
with a focus on design simplicity and strict quality silica column is polished flat to ensure a clean o
columns are specially designed for high-resolution l
control. As a result, they deliver outstanding connection to the emitter of choice. PicoFrit u
analyses of tryptic, natural, and synthetic peptides. m
chromatographic performance on any nano columns eliminate post-column performance
The columns are often applied for LC-MS/MS
n
LC system. losses by spraying directly from the column,
peptide mapping for protein identification, s
biomarker discovery,and systems biology. Due The BioBasic KAPPA line meets all the sensitivity boosting MS performance compared to that a
to their high loading capacity, the columns are needs of demanding LC/MS separations. High provided by a column attached to a tip. n
d
exceptionally suitable for the analysis of low efficiency capillaries are available in internal

abundant peptides in complex proteomics samples. diameters ranging from 500μm all the way down A
Columns for Carbohydrates c
Acclaim PepMap Trap columns are typically to 75μm ID, and lengths of 50mm to 250mm.
HyperREZ XP Carbohydrate columns are based c
applied for the desalting and preconcentration The BioBasic KAPPA line is ideal for all LC/MS e
of peptides before LC separation with MS analyses, especially proteomics separations of on a monodisperse resin with a 4 or 8% s
divinylbenzene content, and provide an ideal s
detection. The columns are designed to provide typically small sample concentrations. BioBasic o
medium for the analysis of carbohydrates and
the highest efficiencies for one dimensional 18, 8, 4 and SCX columns are also available ri
organic acids. Unlike silica based columns they e
peptide mapping experiments and 2D-LC analyses. in nanobore formats for nanospray LC/MS s
are stable at low pH, allowing the use of dilute
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oLCf p crooutepilnesd a tnod M peSp.tides using nano and capillary onfofiesre hriagthioe rt hsaenn stirtaivdiittyio wnaitlh e glerectartoesrp sriagyn. al-to- C
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Bio Columns Selection Guide s
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Analyte Mode of analysis Recommended column io
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BioBasic SEC 4-135 o
Size Exclusion MAbPac SEC-1 4-131 le
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BioBasic AX 4-136 u
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ProPac SCX-10, WCX-10, SAX-10, WAX-10 4-121 s
Ion Exchange
MAbPac SCX-10 4-130
ProSwift IEX 4-127
Proteins
BioBasic 18, 8, 4 4-138
Reversed Phase Acclaim 300 C18 4-141
ProSwift RP 4-128
Hydrophobic Interaction ProPac HIC-10 4-125
ProPac IMAC-10 4-126
Affinity
ProSwift ConA-1S 4-129
Acclaim PepMap 4-142
PepSwift 4-145
Proteomics EASY-Column 4-146
PicoFrit, IntegraFrit 4-147
Peptides KAPPA 4-148
BioBasic 18, 8, 4 4-138
Analytical
Acclaim 300 4-056
Preparative BioBasic Inquire
Amino Acids Ion Exchange AminoPac PA10 Inquire
(derivatized) Reversed phase Hypersil GOLD 4-056
Amino Acids Ion Exchange AminoPac PA10 Inquire
(underivatized) Reversed phase Hypercarb 4-097
Anion Exchange BioBasic AX 4-136
Nucleotides
Polar retention Hypercarb 4-097
BioBasic AX 4-136
Oligonucleotides Ion Exchange DNAPac PA100, PA200 4-132
DNASwift 4-134
Ligand Exchange HyperREZ XP 4-153
Ion Exchange CarboPac Inquire
Acclaim HILIC 4-079
Carbohydrates
HILIC Hypersil GOLD HILIC 4-070
Syncronis HILIC 4-093
Polar retention Hypercarb 4-097
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s HPLC Phases for Biomolecules
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o Particle Size Pore Nominal Surface % USP Phase
s Phase Particle Type (μm) Size (Å) Area (m2/g) Carbon Endcapping Code Code Page
s
e Acclaim PepMap
c 100 C18 Spherical, fully porous silica 2, 3, 5 100 300 15 Yes L1 – 4-142
c
A 300 C18 Spherical, fully porous silica 5 300 100 9 Yes L1 – 4-143
100 C8 Spherical, fully porous silica 3, 5 100 300 9 Yes L7 – 4-144
d
n 300 C4 Spherical, fully porous silica 5 300 300 3 Yes L26 – 4-144
a BioBasic

s 18 Spherical, fully porous silica 5 300 100 9 Yes L1 721 4-138
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8 Spherical, fully porous silica 5 300 100 5 Yes L7 722 4-139
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4 Spherical, fully porous silica 5 300 100 4 Yes L26 723 4-140
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l AX Spherical, fully porous silica 5 300 100 3 No – 731 4-136
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C SCX Spherical, fully porous silica 5 300 100 3 No L52 733 4-137

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Columns for Protein Separations
Silica Size Exclusion Chromatography Phases
Particle Size Pore Exclusion limit USP Phase
Phase SEC Type Particle Type (μm) Size (Å) operating range (kDa) Code Code Page
BioBasic
SEC 60 Aqueous Spherical, fully porous silica 5 60 0.1 - 6 – 733 4-135
SEC 120 Aqueous Spherical, fully porous silica 5 120 0.1 - 50 L33 734 4-135
SEC 300 Aqueous Spherical, fully porous silica 5 300 1 - 500 L33, L59 735 4-135
SEC 1000 Aqueous Spherical, fully porous silica 5 1000 20 - 4000 L33 736 4-135
MAbPac
SEC-1 Aqueous Spherical, fully porous silica 5 300 1 - 500 L33, L59 4-131
Silica Hydrophobic Interaction Chromatography Phases
Base Matrix Particle Pore Nominal Surface Solvent pH
Column Phase Target Applications Capacity
Material Size (μm) Size (Å) Area (m2/g) Compatibility Range
ProPac Hydrophobic High resolution Spherical, porous 5 300 100 340mg lysozyme 2M Ammonium 2.5-7.5
HIC-10 Interaction separations of ultrapure silica per 7.8 x 75mm sulfate/ phosphate
13 proteins and with amide/ethyl column salts, organic
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2-2 protein variants surface chemistry solvent for cleanup
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Polymeric Ion Exchange, Reversed Phase and Affi nity Columns L
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Target Base Matrix Functional Recommended Solvent Maximum pH
Column Phase Capacity C
Applications Material Groups Flow Rate Compatibility Backpressure Range
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MAbPac Strong High Resolution Highly crosslinked Sulfonic 30μg/mL 0.2-2.0 50% 3000psi 2.0-12
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SCX-10 Cation separation of divinlybenzene mL/min acetonitrile (21 MPa) u
Exchange monoclonal 10μm nonporous m
antibody variants particles
n
ProPac Weak High resolution Ethylvinylbenzene Carboxylate 6mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12 s
WCX-10 Cation separations of cross linked with lysozyme mL/min acetone. (21 MPa)
a
Exchange proteins and protein 55% divinlybenzene Incompatable n
variants 10μm nonporous with alcohols d
particles and MeOH

A
ProPac Strong High resolution Ethylvinylbenzene Sulfonate 3mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12
c
SCX-10 Cation separations of cross linked with lysozyme mL/min acetone, (21 MPa)
c
Exchange proteins and protein 55% divinlybenzene MeOH
e
variants 10μm nonporous
s
particles s
ProPac Strong High Resolution Divinlybenzene Sulfonic 30μg/mL 0.2-2.0 50% 3000psi 2.0-12 o
SCX-20 Cation separations of 10μm nonporous mL/min acetonitrile (21 MPa) r
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Exchange proteins and protein particles e
variants s
ProPac Weak Anion High resolution Ethylvinylbenzene Tertiary 5mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12 >
WAX-10 Exchange separations of cross linked with amine BSA mL/min acetone, (21 MPa) >
pvaroritaenintss and protein 5105%μm d invionnlypboernozuesn e MeOH, C
particles o
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ProPac Strong High resolution Ethylvinylbenzene Quaternary 15mg/mL 0.2-2.0 80% ACN, 3000psi 2.0-12 m
SAX-10 Anion separations of cross linked with ammonium BSA mL/min acetone, (21 MPa) n
Exchange proteins and protein 55% divinlybenzene MeOH s f
variants 10μm nonporous o
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particles B
ProSwift Reversed Fast protein analysis Monolith; Phenyl 5.5mg/mL 2.0-4 .0 Most common 2800psi 1.0-14 io
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RP-1S Phase with high resolution polystyrene- Insulin mL/min organic (19.2 Mpa)
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divinylbenzene solvents le
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ProSwift Reversed Fast protein analysis Monolith; Phenyl 1.0mg/mL 1.0-10 Most common 2800psi 1.0-14 u
RP-2H Phase with high resolution polystyrene- Lysozyme mL/min organic (19.3 Mpa) le
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divinylbenzene solvents
ProSwift Reversed Fast protein analysis Monolith; Phenyl 0.5mg/mL 1.0-16 Most common 2800psi 1.0-14
RP-3U Phase with high resolution polystyrene- Lysozyme mL/min organic (19.3 Mpa)
divinylbenzene solvents
ProSwift Reversed Fast protein analysis Monolith; Phenyl 2.3mg/mL 0.1-0.3 Most common 1500psi 1.0-14
RP-4H Phase with high resolution polystyrene- Lysozyme mL/min organic
divinylbenzene Solvents
ProSwift Strong Fast protein analysis Monolith; Quaternary 18mg/mL 0.5-1.5 Most common 1000psi 2.0-12
SAX-1S Anion with high resolution polymethacrylate amine BSA (4.6mm) organic (4.6mm)
Exchange solvents 2000psi
(1.0mm)
ProSwift Strong Fast protein analysis Monolith; Sulfonic acid 30mg/mL 0.5-1.5 Most common 1000psi 2.0-12
SCX-1S Cation with high resolution polymethacrylate Lysozyme mL/min organic (4.6mm)
Exchange (4.6mm) solvents
ProSwift Weak Anion Fast protein analysis Monolith; Tertiary 18mg/mL 0.5-1.5 Most common 1000psi 2.0-12
WAX-1S Exchange with high resolution polymethacrylate amine BSA mL/min organic (4.6mm)
(DEAE) (4.6mm) solvents 2000psi
(1.0mm)
ProSwift Weak Fast protein analysis Monolith; Carboxylic 23mg/mL 0.5-1.5mL/min Most common 1000psi 2.0-12
WCX-1S Cation with high resolution polymethacrylate acid Lysozyme (4.6mm), organic (4.6mm)
Exchange 0.05-0.20 solvents 2000psi
(1.0mm)
ProPac Immobilized High resolution Polystyrene Poly (IDA) >60mg 1.0mL/min EtOH, urea, 3000psi 2.0-12
IMAC-10 Metal separation of divinylbenzene grafts lysozyme/ NaCl, non-ionic (21MPa)
Affi nity certain metal- 10μm nonporous mL gel detergents,
binding proteins particles (4 x 250mm) glycerol,
and peptides acetic acid,
guanidine HCl
ProSwift Affi nity Concanavalin A Monolith; Concanavalin 12-16mg 0-1.0mL/min Up to 10% 2000psi 5.0-8
ConA-1S binding glycans, polymethacrylate A ligands of protein methanol
glycopeptides and
proteins
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s Columns for Carbohydrate Separations
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o Particle Size Pore Nominal Surface % USP Phase
s Phase Particle Type (μm) Size (Å) Area (m2/g) Carbon Endcapping Code Code Page
s
HyperREZ XP
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c Carbohydrate H+ spherical, polymer 8.0 – – – – L17 690 4-152
c Carbohydrate Pb2+ spherical, polymer 8.0 – – – – L34 691 4-152
A
Carbohydrate Ca2+ spherical, polymer 8.0 – – – – L19 692 4-152

d
Carbohydrate Na+ spherical, polymer 10.0 – – – – – 693 4-152
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a Organic Acid spherical, polymer 8.0 – – – – L17 696 4-152
s Sugar Alcohol spherical, polymer 8.0 – – – – L19 697 4-152
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o Columns for Oligonucleotide Separations
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L Column Target Applications Base Matrix Material Substrate Crosslinking Latex Crosslinking Capacity Recommended Eluents Recommended Flow Rate Solvent Compatibility Maximum Backpressure pH Range
DNAPac High resolution 13μm diameter 55% 5% 40μeq Hydroxide 1.5mL/min 0-100% 4000psi 2-12.5
PA100 separations of nonporous substrate (28MPa)
single and double agglomerated with
stranded DNA or alkyl quaternary
RNA ammonium
oligonucleotides functionalized latex
100nm MicroBeads
DNAPac High resolution 8μm diameter 55% 5% 40μeq Hydroxide, 1.2mL/min 0-100% 4000psi 2-12.5
PA200 separations of nonporous substrate acetate/ (28MPa)
single and double agglomerated with hydroxide
stranded DNA or alkyl quaternary
RNA ammonium
oligonucleotides functionalized latex
130nm MicroBeads
DNASwift High resolution Monolith; N/A N/A 50mg, NaClO 0.5-2.5mL Most 1500psi 6.0-12.4
4
separations for polymethacrylate of a 20 mer and NaCl Common
purifi cation of substrate agglomerated oligonucleotide Organic
oligonucleotides with quaternary amine Solvents
functionalized latex
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ProPac SCX-10 and WCX-10 L
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Strong and weak cation exchange columns are based on a nonporous core particle C
providing exceptionally high resolution and efficiency for separations of protein variants o
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(cid:116) Characterization and quality control assessment of monoclonal antibodies and other proteins n
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(cid:116) Unequalled high resolution separations a
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(cid:116) High-effi ciency peaks
d
(cid:116) Unmatched column-to-column and lot-to-lot reproducibility
A
(cid:116) Useful for characterization of related protein variants including deamidation and MAb lysine c
c
truncation variants
e
(cid:116) ProPac SCX-10 is a strong cation exchange column with sulfonate functional groups s
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(cid:116) ProPac WCX-10 is a weak cation exchanger with a carboxylate functional groups o
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control and fast mass transfer for high-efficiency separation and moderate capacity.
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ProPac WCX-10 Ion Exchange HPLC Columns m
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Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID 9.0mm ID 22.0mm ID s
f
10.0 50 – 074600 – – o
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100 – SP5829 – – B
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150 – SP6703 – – m
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250 063472 054993 063474 SP5482 le
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ProPac WCX-10 Ion Exchange HPLC Guards le
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Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID
10.0 50 063480 054994
ProPac SCX-10 Ion Exchange HPLC Columns
Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID 9.0mm ID 22.0mm ID
10.0 250 063456 054995 063700 SP5522
ProPac SCX-10 Ion Exchange HPLC Guards
Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID
10.0 50 063462 054996
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s MAb separation on ProPac WCX-10 column: characterization of
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lysine truncation variants
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o Column: ProPac WCX-10 (4 × 250mm)
s Lysine truncation Eluents: (E1) 20mM MES
s variants* + 115mM NaCl
e + 1mM EDTA, pH 5.5
c K (E2) 20mM MES
+ 145mM NaCl
c
A KK Gradient: t (+m 1inm) M %EDET1A , pH% 5E.25
U 0 100 0
d A MAb sample as received 2 100 0
n 40 0 100
a 60 0 100
s Acvidairci acnhtasrge Bavsaicr icahnatsrge SFlaomwp Rlea:t e: M1.0AmbL P/DmLin
n (cid:3): 280nm
m * Peak assignment supported by data from R.J. Harris,
et.al, J.Chromatogr., A 1995,705, 129–134. and
After digestion with carboxypeptidase B
u Carboxypeptidase B digest.
0 10 20 30 40
ol Minutes 21104
C

C Separation of hemoglobin variants
L
Column: ProPac SCX-10, 4 × 250mm
1.60 1 Eluent: A) 20mM Sodium phosphate,
4mM Potassium cyanide, pH 6
B) 1 M Sodium chloride in water
C) Water
2
Gradient: Time %A %B %C
AU 3 4 Init 50 0 50
30 min 50 50 0
Flow Rate: 1mL/min
Inj. Vol.: 10μL
Detection: 220nm
0 Sample: 1. Fetal hemoglobin
0 5 10 15 20 25 2. Hemoglobin
Minutes 14067 3. Sickle cell hemoglobin
4. Hemoglobin C
ProPac Kits
Part number Phase Description Set Contents Column dimensions
SP5731 ProPac WAX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm
SP5732 ProPac WAX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm
SP5729 ProPac SAX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm
SP5730 ProPac SAX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm
SP5512 ProPac WCX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm
13 SP5513 ProPac WCX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm
0
2
2- SP5727 ProPac SCX-10 Lot Select Column Set 3 columns from 1 lot of resin 250 x 4mm
1
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2s SP5728 ProPac SCX-10 Lot Select Column Set 3 lots of resin, 1 column from each lot 250 x 4mm
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ProPac SAX-10 and WAX-10 L
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Strong and weak anion exchange columns are based on a nonporous core particle providing C
unequalled high resolution and efficiency in the separations of protein variants o
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(cid:116) Unequalled resolution n
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(cid:116) High-effi ciency peaks
a
(cid:116) Useful for characterization and quality control assessment of closely-related protein variants n
d
(cid:116) Supports separation of proteins that differ by as little as one amino acid residue
A
(cid:116) Neutral hydrophilic coat that eliminates protein-resin hydrophobic interactions c
c
(cid:116) Superior lot-to-lot and column-to-column reproducibility e
s
(cid:116) ProPac SAX-10 is a strong anion exchange column with quaternary amine functional group
s
(cid:116) ProPac WAX-10 column is a weak anion exchanger with a tertiary amine functional group o
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transfer for high peak efficiency and resolution separations. C
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ProPac WAX-10 Ion Exchange HPLC Columns m
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Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID 9.0mm ID 22.0mm ID s f
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10.0 250 063464 054999 063707 SP5598 r
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ProPac WAX-10 Ion Exchange HPLC Guards o
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Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID u
10.0 50 063470 055150 le
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ProPac SAX-10 Ion Exchange HPLC Columns
Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID 9.0mm ID 22.0mm ID
10.0 250 063448 054997 063703 SP5594
ProPac SAX-10 Ion Exchange HPLC Guards
Particle Size (μm) Length (mm) 2.0mm ID 4.0mm ID
10.0 50 063454 054998
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s Resolution of phosphorylation variants of ovalbumin
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r Column: ProPac SAX-10, 4 x 250mm
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Eluents: A) Water
s B) 2.0 M NaCl
s C) 0.1 M Tris/HCl (pH 8.5)
e Gradient: Time %A %B %C
c U Init 80 0 20
c mA 15 min 67.5 12.5 20
A Flow Rate: 1.0mL/min
Inj.Volume: 30μL
d Ovalbumin + alk. phos. Detection: 214nm
Sample: Ovalbumin before and after
n Ovalbumin alkaline phosphatase treatment
a
0 2 4 6 8 10 12 14 16
s
Minutes 16445
n
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Effect of sialylation on transferrin chromatography
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ol Columns: ProPac SAX-10, 4 × 250mm
C Eluents: A) Water
B) 2.0 M NaCl
C C) 0.2 M Tris/HCl (pH 9)
HOLO Gradient: Time %A %B %C
L U Init 87 3 10
mA HOLO + neur 30 min 83 7 10
Flow Rate: 1.0mL/min
APO Lot #1 Inj. Volume: 50μL
APO Lot #1+neur Detection: 214nm
Samples: HOLO (iron rich) and APO (iron poor)
human transferrin samples before and after
neuraminidase treatment. Digestions were carried out
overnight at 37 C in sodium acetate buffer at pH 5.
APO Lot #2
APO Lot #2 + neur
0 4 8 12 16 20 24 28 32
ProPac PA1
For hydrophilic anionic protein separations
(cid:116) Good for hydrophilic anionic proteins and peptides
(cid:116) Ideal for high-resolution separations of proteins with pI values from 3 to 11
(cid:116) Available in semipreparative format
(cid:116) Pellicular packing ensures high-effi ciency and fast mass transport
The ProPac PA1 column supports the analysis and purification of hydrophilic anionic proteins and peptides.
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20 ProPac PA1 Ion Exchange HPLC Columns
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u 10.0 50 039657 –
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Co 250 039658 040137
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mn Gradient separation of protein standards
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gra 2 3,4 5 Eluent: 10 to 350mM NaCl
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at in 1.0mM Tris, pH 8.2
m Flow Rate: 5mL/min
hro U 1 6 Detection: UV, 280nm
c C mA Peaks: 1. Horseheart Myoglobin 33μg
fi 2. Contaminant –
nti 3,4. Conalbumin 66
cie 5. Ovalbumin 66
S 6. Soybean Trysin Inhibitor 66
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Seconds 3828
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ProPac HIC-10 L
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Hydrophobic Interaction Chromatography columns for the high resolution separation C
of proteins and peptides o
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(cid:116) High-resolution HPLC separation of proteins, protein variants and peptides n
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(cid:116) Proteins are separated under non-denaturing conditions
a
(cid:116) High protein loading capacity for protein purifi cation applications n
d
(cid:116) Wide range of applications
A
(cid:116) Based on 5μm ultrahigh purity spherical silica gel particles with 300Å pores c
c
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The ProPac HIC-10 column is a high-resolution, high-capacity, silica-based HIC column that provides
s
excellent separations of proteins and variants for analytical and preparative applications. ProPac HIC o
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columns provide exceptional hydrolytic stability under the highly aqueous conditions used in HIC. i
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ProPac PA1 Ion Exchange HPLC Columns >
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Particle Size (μm) Length (mm) 2.1mm ID 4.6mm ID 7.8mm ID C
10.0 75 – – 063665 o
100 063653 063655 – lum
250 – 074197 – n
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600 Column: 7.5 × 75mm (Competitors A and B); Separation of a mixture of m
7.8 × 75mm (ProPac HIC-10) o
4 100 Flow: 1.0mL/min proteins using the ProPac lec
mAU 1 2 3 5 WE luVeLn: ts: 2((AB1))4 20n .Mm1 M(N HN4a)2H SPOO4 in ( 0p.H1 M7. 0N)aH2PO4 (pH 7.0) HseICpa-1ra0t, icoonm opna trwedo tcoo mthpee staitmore ules
2 4
Sample: Mixture of proteins
0 (1mg/mL each fi nal after 1:1 dilution with eluent A) columns
Inj. Volume: 20μL
Competitor A Order of elution: 1. Cytochrome c
Competitor B 2. Myoglobin
-300 3. Ribonuclease A
0 10 20 30 40 50 60 70 4. Lysozyme
Minutes 21286 5. Chymotrypsinogen
1000
10% CH3CN in eluent B Column: ProPac® HIC-10, 4.6 × 100mm Gradient separation of a
100 Eluents: (A) 0.5 M (NH) SO
42 4 monoclonal antibody using the
in 0.1M NaHPO (pH 7.0)
2 4
70 (B) 0.1 M NaH2PO4 (pH 7.0) ProPac HIC-10
Gradient: 70–100% B in 15 min
U
A Flow: 1mL/min
m
Inj. Volume: 5μL (25μg)
Detection: UV, 214nm
%B
Sample: MAb 50μL (50mg/mL)
+ 450μL Eluent B
0
-100
0 2.0 4.0 6.0 8.0 10.0 12.2
Minutes 21538
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s ProPac IMAC-10
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or IMAC column for analytical and semipreparative protein and peptide applications
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c (cid:116) High-purity separations of metal-binding proteins
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A (cid:116) State-of-the-art technology for reusable columns with metal tailored specifi city

d (cid:116) Resolve target proteins using a single column in a high-resolution gradient run
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a (cid:116) Retention control by imidazole concentration or pH gradient
s (cid:116) High loading capacity for protein purifi cation applications
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m (cid:116) Wide range of metal-specifi c applications
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o The ProPac IMAC-10 is a high-resolution analytical and semipreparative column for separation of proteins
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and peptides by immobilized metal affinity chromatography

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ProPac IMAC-10 Immobilized Metal Affi nity HPLC Columns
Particle Size (μm) Length (mm) 1.0mm ID 2.0mm ID 4.0mm ID 9.0mm ID 22.0mm ID
10.0 50 063617 063272 063276 063615 –
250 – 063274 063278 063280 063282
Accssesories for the ProPac IMAC-10 column
Description Cat.No.
IMAC Loading Column (4 x 50mm) 063667
IMAC Loading Column (9 x 50mm) 063710
IMAC Loading Column (9 x 250mm) 063718
Separation of standard proteins with surface-exposed histidines
0.020
2 Column: ProPac IMAC-10 (4mm x 250mm)
1 3 E1: 20mM MES + 0.5 M NaCl, pH = 7.5
Proteins with surface exposed histidine E2: E1 + 100mM imidazole, pH = 7.5
U Gradient: t (min) %E1
mA 0 96
15 0 curve 7
40 0
Inject Vol: 15μL
013 -00..000002 ribonuclease A myoglobin S ample: 12.. 01..0250mmgg//mmLL rmibyoongulocbleinase A
2-2 0 5 10 15 20 25 30 35 40 3. 1.50mg/mL carbonic anhydrase
01 Minutes 20416 Flow Rate: 0.5mL/min
2s Wavelength: 280nm
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a
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su Separation of prion-related peptides using the ProPac IMAC-10
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s and 0.070 1 2 34 7 CElouelunmt: n: ProPac I(EM1)A 20Cm-1M0 H (E4P ×E S2 +5 00.m5 Mm N)aCl, pH = 7.5
mn (E2) E1 + 50mM imidazole, pH = 7.5
u Gradient: t (min) %A
Col 0 95
y 10 0 curve 7
h
p 30 0
gra AU 5 1 2 3 4 5 6 7 Inj. Volume: 15μL
ato m Sample: Prion-derived peptides
m 6 Flow Rate: 0.5mL/min
hro 5i Wavelength: 280nm
fi c C 3i 4i 6i P1 e ak #P HSGeGquGeWncGeQ
mo Scienti -00..00000701 rep5eat2 rep1e0ats4 re1p5eats20 25 30 5234 KPKKHKKKGRRRPPPXKK(GPPPWH(WPGGHGXQGGQGW(PGHGWGQGG)2QG)W2GQ)4
her Minutes 20420 67 WWGGQA((PPHHGGGGGGWWGGQA))4
T 4
X = sarcosine (N-methyglycine)
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