HiTrap Heparin HP columns are prepacked with Heparin Sepharose High Performance for high-resolution purification of proteins with affnity for heparin such as DNA-binding proteins, coagulation factors, lipoproteins, and protein synthesis factors.

Heparin binds to a wide range of biomolecules via both affinity and ion exchange mechanisms.
Immobilized heparin is used for purification of antithrombin III, coagulation factors, and other plasma proteins.
Simple operation with a syringe, a pump or a chromatography system such as ÄKTA design
Stable over pH 5 to 10 and tolerates all commonly used aqueous buffers.

Immobilized heparin has two main modes of interaction with proteins. It can operate as an affinity ligand with affinity for a wide range of biomolecules, including coagulation factors and other plasma proteins, lipoproteins, protein synthesis factors, enzymes that act on nucleic acids, and steroid receptors. Heparin also functions as a high capacity cation exchanger, due to its anionic sulfate groups. Gradient elution with salt is most commonly used in both cases.

Heparin is coupled to the Sepharose High Performance base matrix via the N-hydroxysuccinamide coupling method, to give high capacity, performance, and low leakage levels. HiTrap Heparin HP is delivered in 0.2 M NaCl with 20% ethanol as bacteriostatic agent. Store HiTrap Heparin HP columns equilibrated with 25 ml of 20% ethanol or a buffer containing a bacteriostatic agent. Equilibration is easily performed with a syringe and the supplied Luer adapter. Seal the column with the supplied stopper and domed nut. The recommended storage temperature is 4°C to 30°C.

HiTrap Heparin HP — Technical Specifications

Bed Volume	1 ml
Bed Dimensions	7 x 25 mm
Flow rate	<4 ml/min¹⁾
Storage Conditions	4 to 30°C, 20% Ethanol
Pressure Max. [Over the Packed Bed During Operation]	3 bar [0.3 MPa] (42 psi)
¹⁾The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.

Media

Ligand	Porcine heparin
Average Particle Size	34 μm
Matrix	Highly cross-linked agarose, 6%
Ligand Concentration	10 mg heparin/ml medium
pH stability Working Range	5-10
pH stability Cleaning	5-10
Storage Conditions	4 to 30°C, 20% Ethanol
Chemical Stability	All commonly used buffers

Column

Complete Packsize	1 ml
Column i.d.	7 mm
Material [Column Hardware]	Polypropylene (PP)

Animation
HiTrap: Connecting to a chromatography system (ÄKTA) Close
HiTrap: To use with a syringe Close
Protein purification strategy (Capture, Intermediate Purification, Polishing) Close
The principle of Affinity Chromatography Close

Application note
Partial purification of DNA binding proteins using HiTrap Heparin Summary This work describes partial purification of three different DNA binding proteins, i.e. H-NS, RNA polymerase and Oct-1, using pre-packed HiTrap Heparin 5 ml columns in the initial chromatographic step. H-NS was purified directly from a bacterial lysate using heparin affinity chromatography and gel filtration as the only chromatographic steps. The protein was shown to be 70% pure by SDS-PAGE. RNA polymerase was purified using gel filtration and ion exchange chromatography after the heparin step. SDS-PAGE analysis showed the polymerase to be 90% pure. Recombinant human Oct-1 was shown to bind to HiTrap Heparin, but was not further purified. The results show that HiTrap Heparin is a suitable column for the binding of both native and recombinant DNA binding proteins directly from bacterial lysates. Close

Application note

Partial purification of DNA binding proteins using HiTrap Heparin Summary

Data file
HiTrap Heparin HP Summary HiTrap™ Heparin HP is one of a range of prepacked, ready to use columns for affinity chromatography. Fast, simple and easy separations are provided by the prepacked column with a high performance affinity medium. HiTrap Heparin HP is particulary suitable for the isolation and purification of antithrombin III and other coagulation factors, lipoproteins, lipases, protein synthesis factors, hormones, steroid receptors, nucleic acid binding enzymes and interferon. • Fast and convenient to use • Packed with Heparin Sepharose™High Performance • Simple operation with a syringe, a pump or a chromatographic system such as ÄKTAdesign™ Close

Data file

HiTrap Heparin HP Summary

Handbook
Affinity Chromatography, Principles & Methods Summary This handbook describes the role of affinity chromatography in the purification of biomolecules, the principle of the technique, the media available and how to select them, application examples and detailed instructions for the most commonly performed procedures. Practical information is given as a guide towards obtaining the best results. Close
Strategies for protein purification Summary Strategies for Protein Purification is focused on chromatographic purification of proteins at research scale. Beginners can use the handbook to get a quick start when entering the field; experienced protein purifiers may also find valuable information. The handbook describes how to design a protein purification procedure, how to select and combine chromatography methods and purification formats, and how to monitor and evaluate protein purification. It gives an overview of the available methods and provides advice on how to avoid pitfalls in all operations, from initial sample preparation to final analysis of the purified protein. Close

Handbook

Affinity Chromatography, Principles & Methods Summary

Strategies for protein purification Summary

Instruction/protocol
HiTrap Heparin HP, 1 ml and 5 ml Summary HiTrap™ Heparin HP is a prepacked ready to use, column for preparative affinity chromatography. The special design of the column, together with the matrix, provide fast, simple and easy separations in a convenient format. The columns can be operated with a syringe, peristaltic pump or liquid chromatography system such as ÄKTAdesign™ or FPLC™ System. Close

Instruction/protocol

HiTrap Heparin HP, 1 ml and 5 ml Summary

Magazine article
Redesigned HiTrap fittings offer easier connection to ÄKTAdesign chromatography systems (Life Science News 18, p19) Summary The new inlet and outlet design of HiTrap columns eliminates the need for column connectors when used with ÄKTAdesign™ chromatography systems. Connection to a syringe, laboratory pump, or chromatography system such as FPLC™ System is also possible using the provided connectors. The guide on this page shows the different types of connector and explains their use. Close

Magazine article

Redesigned HiTrap fittings offer easier connection to ÄKTAdesign chromatography systems (Life Science News 18, p19) Summary

Material Safety Data Sheet
HiTrap Heparin HP 1 ml, 5 x 1 ml Close
HiTrap Heparin HP 1 ml, 5 x 1 ml Close

Poster
HiTrap Heparin HP a Useful Tool in Designing Large Scale Purification on Heparin Sepharose 6 Fast Flow Summary Affinity chromatography using immobilized heparin is an important way of purifying Antithrombin III (blood coagulation protein) both from plasma and from recombinant sources. Amersham Biosciences has developed two affinity media which contain immobilized heparin: 1 ml and 5 ml prepacked HiTrap™ columns useful for small scale screening work and a gel for large scale process use, Heparin Sepharose™ 6 Fast Flow. This poster presents general properties of the two products and a methodological study illustrating how HiTrap columns can be used for fast small scale optimization of the purification process. It will also be shown how the optimized process can be directly scaled-up from High Performance to Fast Flow gel. Close

Poster

HiTrap Heparin HP a Useful Tool in Designing Large Scale Purification on Heparin Sepharose 6 Fast Flow Summary

Reference list
HiTrap Heparin HP Summary The reference list for this product. Close

Selection guide
Affinity chromatography columns and media Summary This Selection Guide for affinity chromatography columns will help you find the right column or media. Areas: 1) group specific purification 2) isolation and purification of immunoglobulins 3) recombinant fusion proteins 4) coupling ligands 5) metal chelate chromatography Close
Prepacked chromatography columns for ÄKTA™ design systems Summary Selection guide describing all the prepacked columns available from GE Healthcare for use with ÄKTA™ chromatography systems. Close
Protein and Peptide Purification: Technique selection guide Summary This technique selection guide is designed to help you in selecting the technique most suited to your application. It covers ion exchange chromatography (IEX), reversed phase chromatography (RPC), hydrophobic interaction chromatography (HIC), gel filtration (GF), affinity chromatography (AC). Close
Protein sample preparation Selection Guide Summary Almost all protein samples need further preparation after collection. The quality of sample preparation is crucial to get the best possible analytical results. This selection guide helps you to find the appropriate methods and products to obtain proteins for further studies in scales from nanograms to milligrams of purified protein. Three separate workflows are available for different sources of target protein; recombinant proteins, antibodies, and proteins from natural sources. These workflows are further refined and adapted to suit different number of samples, from a few to many such as in screening applications. Close

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Accessories

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Running HiTrap, HisTrap and GSTrap with a stand alone pump

Running HiTrap, HisTrap and GSTrap with a syringe

Running HiTrap, HisTrap and GSTrap with ÄKTAdesign systems

For accessories not displayed in this section, please contact your local GE Healthcare sales representative.

Running HiTrap, HisTrap and GSTrap with a stand alone pump

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#	Product Name	Product Code	Price
1	Tubing Cutter	18-1112-46		Price on request
1	Tubing, 2 m, i.d. 0.75 mm, o.d. 1/16"	18-1112-53	CAD 73.19	Add
1	Tubing i.d. 0.5 mm, o.d. 1/16"	18-1113-68	CAD 48.33	Add
1	Tubing, 2 m, i.d. 1.0 mm, o.d. 1/16"	18-1115-83	CAD 53.32	Add
1	Tubing i.d. 0.25 mm, o.d. 1/16"	18-1120-95	CAD 56.46	Add
2	HiTrap/HiPrep, 1/16", male connector for ÄKTAdesign	28-4010-81	CAD 81.39	Add

Running HiTrap, HisTrap and GSTrap with a syringe

/gehcls_images/GELS/Images/TechSupportImages/Files/18114736_47.jpeg

#	Product Name	Product Code	Price	Qty
1	Connector 1/16" Male/Luer Female	18-1112-51	CAD 82.59		Add

Running HiTrap, HisTrap and GSTrap with ÄKTAdesign systems

/gehcls_images/GELS/Images/TechSupportImages/Files/18114736_45.png

#	Product Name	Product Code	Price
1	Tubing Cutter	18-1112-46		Price on request
1	Tubing, 2 m, i.d. 0.75 mm, o.d. 1/16"	18-1112-53	CAD 73.19	Add
1	Tubing i.d. 0.5 mm, o.d. 1/16"	18-1113-68	CAD 48.33	Add
1	Tubing, 2 m, i.d. 1.0 mm, o.d. 1/16"	18-1115-83	CAD 53.32	Add
1	Tubing i.d. 0.25 mm, o.d. 1/16"	18-1120-95	CAD 56.46	Add
2	HiTrap/HiPrep, 1/16", male connector for ÄKTAdesign	28-4010-81	CAD 81.39	Add
3	Fingertight Connector 1/16" Male for Tubing o.d. 1/16"	18-1112-55	CAD 94.1	Add