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Affinity Resins
96-Well Plates
Spin Columns
Bulk Resins
Purify!
True one-step histidine-tagged protein
purification with Sigma’s HIS-Select Technology ®
■ S
uperior selectivity for higher purity Sigma’s HIS-Select products purify histidine-tagged proteins quickly
of target proteins and with high selectivity. This is due to Sigma’s patented HIS-Select
■ N
on-charged, hydrophilic linkage
metal chelate linker, which is hydrophilic and non-charged. Because
reduces non-specific binding
■ O
ne-step purification
the HIS-Select linkage chemistry is non-charged, nonspecific binding
of unwanted proteins is greatly diminished, allowing for higher purity
of your histidine-tagged protein. Most other immobilized metal
affinity chromatography (IMAC) systems for histidine-tagged protein
purification utilize a charged chelate linkage.
1
2
TM
Low Range SigmaMarker (M3913)
Whole Cell Extract in Sample Buffer
7
8
Elute 1
Elute 2
For more information, visit sigma.com/hisselect
3 Whole Cell Extract with CelLytic™ B 9 Elute 3
4 Flow Through 10 Elute 4
5 Wash 1 11 Low Range SigmaMarker
6 Wash 2 (M3913)
HIS-Select Nickel Affinity Gel (P6611) HIS-Select High Capacity (HC) Nickel Coated Plates (S5563)
This product is the tool of choice for small to medium scale HIS-Select High Capacity (HC) Nickel Coated Plates are coated with a
purification of histidine-tagged recombinant proteins. The unique, proprietary, high-density nickel chelate matrix. This patented coating
non-charged, hydrophilic linkage of the nickel chelate group to the allows for greater per-well binding capacity (≥4 µg/well) than any
agarose ensures high selectivity for histidine-tagged proteins and other commercial histidine-binding plate and still demonstrates low
reduces non-specific binding of other proteins and is compatible with non-specific binding. These plates allow for the in-well quantification
both denaturing or non-denaturing conditions. of bound protein and are suitable for applications including:
Expression Screening, PAGE, MALDI-MS, and High Throughput
HIS-Select HF Nickel Affinity Gel (H0537)
Purification.
HIS-Select HF (High Flow) Nickel Affinity Gel brings the superior
selectivity of HIS-Select Immobilized Metal Affinity Chromatography HIS-Select iLAP® 5 mL column (H9913)
(IMAC) technology to a highly crosslinked agarose designed for higher HIS-Select iLAP (Integrated Lysis and Affinity Purification) columns
flow rates and mechanical stability under pressure. This product is are single-use, disposable columns designed for one-step purification
designed for production scale purification and FPLC™ applications. of histidine-tagged proteins directly from a 5 mL bacterial culture.
As with other HIS-Select products, the non-charged, hydrophilic The single-step method uses our innovative, patent-pending iLAP
linkage of the NTA chelate group to the agarose ensures true one-step technology, which allows quick and simple colony screening for
purification. histidine-tagged recombinant clones. Each column is capable of
purifying at least one milligram of histidine-tagged protein in less than
HIS-Select Cobalt Affinity Gel (H8162)
an hour, resulting in a nearly pure fusion protein preparation following
For those who prefer to use cobalt as the affinity metal ion for
column elution.
histidine-tagged protein purification, this product allows for high
purity, low non-specific binding, and high binding capacity for your HIS-Select Filter Plate (H0413)
protein. This product also works well for purification of histidine- The filter plate is an immobilized metal-ion affinity chromatography
tagged proteins in native, denaturing, or mild reducing conditions. (IMAC) product that incorporates the speed, consistency, and
convenience of spin columns into a multi-sample format. This pre-
HIS-Select Spin Columns (H7787)
packed 96-sample plate has wells that contain 20 µm, spherical silica
Prepacked and ready-to-use HIS-Select Spin Columns allow for fast particles (100 nm pore size), which are derivitized with a nickel chelate
and consistent small-scale purification of histidine-tagged protein linkage. The particles allow for small scale histidine-tagged protein
from crude cell extracts. The Spin Columns contain a matrix of silica purification from crude cell extracts in less than 15 minutes. The filter
particles and use the patented HIS-Select technology to attain high plates are also compatible with centrifugation, vacuum-manifolds,
protein purity and very low non-specific binding. Spin Columns allow and robotics systems.
for fast, one-step purification of at least 150 µg of histidine-tagged
HIS-Select Wash Buffer (H5288)
protein in less than 15 minutes.
The buffer is a prepared solution of 10 mM imidazole (in 0.3 M
EZview™ Red HIS-Select HC Nickel Affinity Gel (E3528) sodium chloride, and 50 mM sodium phosphate, pH 8.0) that is
optimized to reduce non-specific binding of proteins during wash
When performing small scale affinity capture, such as steps when purifying histidine-containing proteins using
immunoprecipitation, the affinity matrix is difficult to see in the HIS-Select purification products or other immobilized metal affinity
microcentrifuge tubes. Accidental aspiration of the resin leads to chromatography (IMAC) products.
quantitative variability in results. The EZview Red Affinity Gel greatly
reduces the risk of pellet loss. EZview resins perform as well as HIS-Select Elution Buffer (H5413)
conventional non-colored affinity gels, but allow the user to easily The buffer is a prepared solution of 250 mM imidazole (in 0.3 M
differentiate pellet from supernatant. This correlates to more accurate sodium chloride, and 50 mM sodium phosphate, pH 8.0) that is
data because less target protein is lost. optimized to elute histidine-containing target proteins from
HIS-Select purification products or other immobilized metal affinity
HIS-Select High Sensitivity (HS) Nickel Coated Plates chromatography (IMAC) products.
(S5688)
HIS-Select High Sensitivity (HS) Nickel Coated Plates are designed
for highly accurate low-level detection of histidine-tagged proteins.
The captured proteins can be detected using standard enzyme-
linked assay (ELISA) techniques and are suitable for high throughput
applications. These plates are manufactured by utilizing a unique
non-charged hydrophilic linkage for the nickel-chelate, are pre-treated
to reduce non-specific binding, and are sensitive enough to capture as
little as 1 ng/well of histidine-tagged protein. The plates are supplied
as 96-well strip plates.
HIS-Select® Quick Reference Guide
Scale per Sample 100 µg–10 g 100 µg–100 g 100 µg–10 g 150 µg 10–500 µg per
600 µl/load pull-down
Binding Capacity 15 mg/mL 15 mg/mL 15 mg/mL 150 µg/column 15 mg/mL
Matrix 6% Beaded Agarose 6% Beaded Agarose, 6% Beaded Agarose Silica Particles 6% Beaded Agarose
Highly Cross-linked (Porous, Spherical)
Bead Size 45–165 µm 45–165 µm 45–165 µm ~20 µm 45–165 µm
E xclusion Limit 4 × 10 Da6
4 × 10 Da6
4 × 10 Da6
0.1 µm 4 × 106 Da
(MW) or Pore Size (~10 × 106 Da)
ax. Linear Flow
M 150 cm/hr 3000 cm/hr 150 cm/hr N/A N/A
Rate (Max. pressure) (5 psi) (200 psi) (5 psi)
ecommended
R 1 mL/min 5 mL/min 1 mL/min N/A N/A
Flow Rate for
1 × 2 cm Column
ecommended
R N/A N/A N/A Equivalent to ~80 × g 1 hour at 4 °C
Binding Time/Speed Centrifugation
ptimal pH
O 3-10 3-10 3-10 3-10 3-10
Stability
Physical Form 50% suspension in 50% suspension in 50% suspension in Prepacked dry matrix in 50% suspension in
30% ethanol 30% ethanol 30% ethanol spin column 30% ethanol
Antimicrobial Agent 30% ethanol 30% ethanol 30% ethanol N/A 30% ethanol
Storage 2-8 °C 2-8 °C 2-8 °C 2-8 °C 2-8 °C
ecommended
R 0-10 mM 0-10 mM 0-10 mM 0-5 mM 0-10 mM
Imidazole Conc.
for Load/Wash
ecommended
R 250 mM 250 mM 250 mM 250 mM 250 mM
Imidazole Conc.
for Elution
High Sensitivity
(HS) High Capacity (HC)
Nickel Coated Strip Nickel Coated HIS-Select iLAP® Nickel Filter Plates HIS-Select HIS-Select
Plates Plates 96-well Column 96-well Wash Buffer Elution Buffer
S5688 S5563 H9913 H0413 H5288 H5413
1 ea., 5 ea. 1 ea., 5 × 1 ea. 1 ea., 25 ea. 1 ea., 5 ea. 500 mL, 1 L 250 mL, 500 mL
n igh Throughput
H n igh Throughput,
H n irect Cell Lysis
D n S mall Scale Optimized to reduce Optimized to elute
Screen (ELISA) High Capacity and Purification of n High Throughput non-specific binding of histidine-containing
n Compatible with Screening Histidine-tagged Screening proteins during wash target proteins from
Robotics n Multi-Sample Protein in a Prepacked n Process by steps when purifying HIS-Select purifica-
Miniprep Column Centrifugation or histidine containing tion products or
n In-well Protein Assays n Small Scale Vacuum proteins with HIS-Select other immobilized metal
n Compatible with n Gravity n Compatible with purification products or affinity chromatography
Robotics Robotics other immobilized metal (IMAC) products
affinity chromatography
(IMAC) products
1 ng–1 µg/well 1 –4 µg/well 100 µg–2 mg 1 mg/well N/A N/A
96-well: 200 µl/well 96-well: 200 µl/well 1 mL/load
Sensitivity: ≤1 ng/well 96-well: ≥4 µg/well 2 mg/column 1 mg/well N/A N/A
Clear Polystyrene Plate; Clear Polystyrene Plate; 6% Beaded Agarose Silica Particles N/A N/A
Proprietary Coating Proprietary Coating Tableted Lysis Reagents (Porous, Spherical)
N/A N/A 45–165 µm ~20 µm N/A N/A
N/A ≥300 kDa 4 × 10 Da6
0.1 µm N/A N/A
(~10 × 106 Da)
N/A N/A N/A N/A N/A N/A
Clear 96-well flat- Clear 96-well flat- Tablets in 5.5 mL Prepacked dry matrix in Liquid Liquid
bottom, bottom, polystyrene polypropylene column 96-well filter plate
polystyrene strip plate plates
N/A chlorhexidine N/A N/A N/A N/A
2-8 °C 2-8 °C 2-8 °C 2-8 °C Room Temperature Room Temperature
0-5 mM 0-1 mM 0-10 mM 0-5 mM N/A N/A
SW-COC-003139
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©2008 Sigma-Aldrich Co. All rights reserved. SIGMA, , SAFC, , SIGMA-ALDRICH, ALDRICH, , FLUKA, , and SUPELCO, are trademarks belonging to Sigma-Aldrich
Technology and Service
Co. and its affiliate Sigma-Aldrich Biotechnology, L.P. iLAP and HIS-Select are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. EZView, SigmaMarker,
and CelLytic are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. FPLC is a trademark of GE Healthcare. Sigma brand products are sold through Sigma-Aldrich,
Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the
product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.
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