All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: PEMAB-1 (for 1ml, 25%(v/v))

Cat. No.: PEMAB-5 (for 5ml, 25%(v/v))

Cat. No.: PEMAB-20 (for 20ml, 25%(v/v))

Description

Phosphoprotein Enrichment Magnetic Agarose Beads are prepared by covalently coupling high‑quality Phospho-tag™ ligands to agarose magnetic beads. Under physiological pH conditions, this product specifically binds phosphoproteins present in samples such as animal, plant, or microbial lysates, serum, or ascites. It is suitable for the purification of phosphoproteins or phosphoprotein complexes, as well as for immunoprecipitation (IP) and co‑immunoprecipitation (Co‑IP) applications.

Phosphorylation is one of the most extensively studied post‑translational modifications. Protein phosphorylation and dephosphorylation play essential roles in numerous biological processes. Mass spectrometry (MS) enables precise identification of phosphorylation sites at the amino acid level and is a core technology in phosphoproteomics. Prior to MS analysis, proteins must be digested into peptides. However, this step increases sample complexity, and the large number of non‑phosphorylated peptides can suppress the MS signals of phosphorylated peptides, reducing detection sensitivity. Therefore, efficient enrichment of phosphoproteins or phosphopeptides prior to MS analysis is critical. Phosphoprotein Enrichment Magnetic Agarose Beads enable rapid and efficient enrichment and purification of phosphoproteins and phosphopeptides. The workflow is simple, the binding conditions are mild, and the purified proteins can be directly used for downstream applications.

This product is developed based on immobilized metal affinity chromatography (IMAC). Studies have shown that the dinuclear zinc(II) complex Phospho-tag™ contains a Zn–Zn vacancy distance of 3.6 Å. Under physiological pH, the oxygen atoms of phosphate groups (ROPO₃²⁻) can coordinate with the fifth coordination site of zinc(II), forming a stable phosphate complex (ROPO₃–Zn₂L³⁺). This property enables selective enrichment and purification of phosphorylated proteins.

Phosphoprotein Enrichment Magnetic Agarose Beads specifically bind phosphoproteins and can be conveniently used with magnetic racks or other magnetic separation devices for phosphoprotein purification, phosphoprotein complex isolation, or immunoprecipitation‑based workflows.

Features

• The product exhibits high stability, strong specificity, and a high binding capacity for target phosphoproteins. It shows excellent selectivity toward phosphorylated proteins. Each milliliter of bead suspension contains approximately 25% agarose magnetic bead slurry and no less than 1 μmol/mL of Phospho-tag™ ligand. Typically, each milliliter of settled agarose magnetic beads can bind more than 4 mg of phosphorylated proteins, with the maximum binding capacity depending on factors such as the molecular weight of the phosphoproteins.
• The product enables rapid binding of target proteins. The Phospho-tag–chelated zinc ions are immobilized on beads with an average particle size of approximately 100 μm. Phosphoprotein adsorption is usually completed within 30 minutes, and purification or immunoprecipitation workflows can be completed within 60 minutes. Shortening the operation time helps prevent degradation or denaturation of target proteins during prolonged handling, thereby preserving protein activity.
• The product allows efficient elution of phosphorylated proteins. Depending on the structural features, biological functions, and downstream application requirements of the phosphoproteins, phosphate-containing buffers are used for multiple elution steps. The elution process is rapid and yields high recovery efficiency.
• The product is easy to use. It is stored in a specially formulated protective buffer that does not contain glycerol. The beads enable rapid and efficient separation through magnetic adsorption, eliminating the need for centrifugation during operation.

Parameters

Each milliliter of product suspension contains approximately 0.25 mL of settled agarose magnetic beads. The labeled volume of this product refers to the total suspension volume. One milliliter of suspension can typically be used to purify approximately 1 mg of phosphorylated protein with a molecular weight of around 60 kDa. The molecular weight of the phosphoprotein will influence the total amount of target protein that can be purified using this product.

Storage

Storage at 4 °C with a shelf life of one year.

Precautions

• Phosphate‑containing buffers should not be used during purification, as phosphate ions may compete with phosphorylated proteins for binding, thereby reducing the binding capacity of the product.
• The pH of all working solutions should be maintained between 6 and 8. Avoid high‑speed centrifugation and drying of the beads. Do not leave the beads in a magnetic field for extended periods, as this may cause bead aggregation.
• During use, the concentrations of buffering reagents such as Tris, HEPES, and MOPS should not exceed 100 mM. The concentrations of non‑ionic detergents such as Triton, Tween, or NP‑40 should not exceed 2%, and the concentrations of sodium deoxycholate or CHAPS should not exceed 1%. Compatibility of other reagents may be inferred from the above guidelines but should be experimentally validated if uncertain.
• Before use, the beads should be fully resuspended by gently inverting the tube several times. Mixing should be gentle; vigorous vortexing is not recommended.
• Positive and negative controls are recommended when performing purification or immunoprecipitation experiments.
• Protein samples should be processed as soon as possible after collection and kept at 4 °C or on ice throughout the procedure to minimize degradation or denaturation. A protease inhibitor cocktail may be added to the sample to inhibit proteolysis, but EDTA should not be used.
• When removing supernatants using a vacuum pump or similar device, the suction force should be carefully controlled to avoid aspirating aggregated beads.
• A low concentration (0.1%) of non‑ionic detergents such as Triton X‑100, Tween‑20, or NP‑40 can effectively prevent bead aggregation without affecting the binding efficiency of the beads.
• When preparing lysis buffers, note that reagents such as DTT, β‑mercaptoethanol, and EDTA may affect the interaction between the product and phosphorylated proteins.
• This product is intended for use by trained professionals in scientific research only. It is not for clinical diagnosis or therapeutic use, not for food or drug applications, and should not be stored in residential environments.
• For your safety, wear a laboratory coat and disposable gloves when handling this product.

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