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Cat. No.: HPAG-1 (for 1ml)

Cat. No.: HPAG-5 (for 5ml)

Cat. No.: HPAG-20 (for 20ml)

Description

Heparin Agarose is a high-quality agarose gel made by covalently coupling heparin with highly cross-linked 6% agarose. Also known as heparin affinity resin, it efficiently separates biomolecules that have an affinity for heparin, including antithrombin III, thrombin, clotting factors, and other plasma proteins. It can also be used to purify lipoproteins, nucleases, steroid receptors, growth factors, interferons, nucleic acid-binding proteins, viruses, and other biomolecules. Heparin agarose gel is widely used for the purification of various proteins and viruses, and the binding, washing, and elution of target proteins can be achieved by altering the salt concentration in the buffer. This product can also be used to remove residual nucleases, including RNase and DNase.

Heparin, commonly found in its sodium salt form (heparin sodium), is a highly sulfated linear polysaccharide primarily composed of repeating units of uronic acid (L-iduronic acid and D-glucuronic acid) and 2-amino-2-deoxy-D-glucose (glucosamine), with an average molecular weight of approximately 15 kDa. Heparin is mainly produced by mast cells and basophils and is abundant in the liver, lungs, blood vessel walls, intestinal mucosa, and mast cells. Its interaction with heparin-binding proteins regulates many important physiological processes, such as complement cascade activity, cell differentiation, growth, migration, inflammation, and pathogen infection.

The interaction between heparin and most proteins primarily depends on the high negative charge density of sulfonic and carboxylic acid groups in heparin, which can form ionic bonds with positively charged basic amino acids (arginine, lysine) in proteins, resulting in strong affinity. Some protein binding to heparin occurs due to hydrogen bonding with polar amino acids (asparagine and glutamine). For example, brain natriuretic peptide (BNP) interacts with heparin primarily through hydrogen bonds. Additionally, the flexible conformation of L-iduronic acid in heparin allows for the formation of specific structural domains that can uniquely bind to proteins, such as those resembling nucleic acids or viral receptors.

Given its biological activity related to anticoagulation, affinity for factor-like proteins, and structural characteristics resembling nucleic acids and viral receptors, heparin agarose gel has extensive applications in protein purification. It can specifically purify antithrombin III, thrombin, clotting factor IX (FIX), clotting factor XI (FXI), apolipoproteins, nucleic acid-binding proteins, transcription factors, nucleases, DNA polymerases, endonucleases, and is also used for the separation and purification of viral vaccines and vectors, as well as growth factors and interferons.

Features

• This product has a high binding capacity. Compared to many similar products, it offers significantly higher binding capacity, allowing for the rapid separation and purification of biomolecules such as antithrombin III, clotting factors, and growth factors from complex samples. The Heparin Agarose in this product is a 50% gel suspension, with each milliliter of Heparin Agarose sediment coupling 4 mg of heparin, capable of binding approximately ≥2 mg of antithrombin III, making it highly effective for experiments such as immunoprecipitation.
• This product has strong specificity. It can specifically bind to antithrombin III, clotting factors, growth factors, apolipoproteins, nucleic acid-binding proteins, and more, resulting in high purity of the obtained products. These can be further used for a series of downstream analytical tests, including Western blotting, ELISA, and mass spectrometry.

This product is a 50% gel suspension, with the total volume containing 0.5 ml of gel sediment per milliliter. If 50 μl of the agarose gel suspension is used for each sample, this product can be utilized for 20 sample reactions per milliliter.

Storage

Store at -20°C, with a shelf life of one year. Store at 4ºC, effective for at least one month.

Precautions

• Before use, this product must be adequately resuspended by gently inverting it several times to ensure the agarose gel is mixed evenly. Avoid vigorous vortexing or shaking to prevent protein denaturation and gel breakdown.
• This product contains trace amounts of preservatives; it is advisable to wash the gel three times with an appropriate solution such as TBS prior to use to eliminate any potential interference from the preservatives.
• When performing immunoprecipitation or purification, it is recommended to set up positive and negative control groups.
• Protein samples should be purified as soon as possible after collection, and they should always be kept at 4ºC or in an ice bath to slow down protein degradation or denaturation. To effectively inhibit protein degradation, an appropriate mixture of protease inhibitors can be added to the protein samples.
• If centrifugation does not completely remove insoluble material from the protein samples, the sample solution can be filtered using a 0.45μm membrane. Agarose that has been eluted with acidic solutions or SDS-PAGE cannot be reused. To minimize heparin loss, the low pH elution step should not exceed 10 minutes, whether done manually or automatically.
• High concentrations of DTT, mercaptoethanol, and guanidine hydrochloride may affect the binding of this product to ligands.
• This product is intended for scientific research use by professionals only and should not be used for clinical diagnosis or treatment, for food or drug purposes, or stored in a regular household.
• For your safety and health, please wear laboratory coats and disposable gloves during handling.

Only for research and not intended for treatment of humans or animals

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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory