Types and Selection of Affinity Chromatography Media
Types and Selection of Affinity Chromatography Media
Affinity chromatography media achieve separation through specific or highly selective interactions between immobilized ligands and target molecules. His-tagged proteins can be captured using Ni²⁺- or Co²⁺-based metal-chelating media, while GST, MBP, and Strep tags correspond to glutathione, Dextrin, and Strep-type ligands, respectively. Boronic acid media can recognize molecules containing cis-diol structures, sulfated media can be used for enrichment of certain proteins and viral particles, and CNBr-activated media are used for custom ligand coupling.
Keywords: affinity chromatography media; metal chelation; His tag; boronic acid affinity; dextran sulfate; activated media; protein purification
1 Technical Basis of Affinity Chromatography Media
1.1 Affinity Recognition Mechanisms
Affinity chromatography relies on reversible and selective interactions between target molecules and immobilized ligands. His tags coordinate with immobilized Ni²⁺ or Co²⁺ through the imidazole groups of histidine residues; GST binds glutathione; MBP binds Dextrin- or Amylose-type polysaccharide ligands; Strep-tag II binds ligands such as Strep-Tactin; and FLAG, HA, c-Myc, V5, and GFP can be captured using corresponding antibodies or nanobodies. In non-tag-based systems, boronic acid groups can form reversible boronate esters with cis-diols, whereas sulfated ligands bind specific biomacromolecules through sulfate-mediated heparin-mimetic interactions and electrostatic interactions.
1.2 Matrix and Ligand Structure
Affinity chromatography media generally consist of a porous matrix, spacer arm, and functional ligand. Agarose matrices have good hydrophilicity and are suitable for purification of proteins and other biomacromolecules. Highly crosslinked Fast Flow matrices have greater mechanical strength and can operate at higher flow rates and column pressures. Pore size affects diffusion of macromolecules into the interior of the medium, ligand density determines the number of potential binding sites, and spacer-arm length and spatial exposure of the ligand directly influence accessibility of the target molecule.
1.3 Binding Capacity and Selectivity
Binding capacity reflects the amount of target molecule that can be captured per unit volume of medium, whereas selectivity determines the degree of separation between contaminants and the target molecule. Higher ligand density can increase theoretical capacity but may also increase nonspecific adsorption. More stringent molecular recognition generally reduces background, but weakly binding targets are more likely to be lost during sample loading or washing. Actual performance can be assessed by examining the distribution of the target protein in the Input, Flow-through, Wash, and Elution fractions.
2 His-Tag and Metal-Chelating Affinity Media
Immobilized Metal Affinity Chromatography (IMAC) immobilizes Ni²⁺ or Co²⁺ through chelating ligands such as IDA, NTA, and carboxymethyl aspartate, after which imidazole groups in the His tag participate in metal coordination to achieve capture. Both the metal ion and the structure of the chelating ligand affect capacity, selectivity, and metal stability.
2.1 Ni-IDA Affinity Media
(1) Binding Characteristics
After IDA immobilizes Ni²⁺, several metal coordination sites remain available for interaction with His tags, and the medium therefore usually provides relatively high binding capacity. His-tagged proteins can bind under neutral to mildly alkaline conditions and can subsequently be eluted using imidazole to compete for metal coordination sites.
(2) Major Advantages
Ni-IDA is suitable for capturing highly expressed His-tagged proteins, particularly for initial purification requiring relatively high medium capacity. Highly crosslinked matrices such as 6FF can further improve flow rate and mechanical stability for processing larger sample volumes.
(3) Major Limitations
When more open metal coordination sites are available, host-cell proteins naturally enriched in residues such as His and Cys may also be adsorbed. Increasing imidazole and salt concentrations in the wash buffer can reduce some background, but excessive imidazole may cause weakly bound target proteins to elute prematurely.
2.2 Ni-NTA Affinity Media
(1) Binding Characteristics
NTA chelates Ni²⁺ more strongly than IDA while retaining metal coordination sites available for His-tag binding. It provides a relatively balanced combination of metal immobilization stability and target-protein capacity and is a commonly used His-tag affinity medium.
(2) Major Advantages
Ni-NTA can be used under native conditions and also in some denaturing systems containing urea or guanidine hydrochloride. Conventional His-tagged proteins can be competitively eluted by increasing imidazole concentration, making the system suitable for scales ranging from expression screening to preparative purification.
(3) Major Limitations
Chelators such as EDTA can compete for immobilized metal ions, and some reducing agents at relatively high concentrations may also affect medium stability. Native metal-binding proteins in the sample may still undergo copurification.
2.3 Co-NTA Affinity Media
(1) Binding Characteristics
Co-NTA uses Co²⁺ as the binding center for His tags. Co²⁺ binds some non-tagged host proteins more weakly and therefore often produces lower nonspecific background.
(2) Major Advantages
Co-NTA is suitable when eluates from Ni²⁺ media still contain substantial background proteins, particularly for low-abundance His-tagged proteins or samples requiring relatively high purity after a single purification step.
(3) Major Limitations
Co²⁺ systems are generally not characterized by maximum binding capacity. When tag exposure is insufficient or target-protein binding to the metal site is weak, flow-through losses may increase.
2.4 Co Carboxymethyl Aspartate Affinity Media
(1) Binding Characteristics
Co carboxymethyl aspartate media use carboxymethyl aspartate-type chelating ligands to immobilize Co²⁺, forming another class of His-tag metal-affinity system. Because the ligand structure differs from NTA, the mode of metal immobilization and target-protein binding characteristics also differ.
(2) Major Advantages
This medium can be used for purification of His-tagged proteins and can also be compared in parallel with Co-NTA, Ni-NTA, and Ni-IDA to examine the effects of different metals and chelating structures on target-protein purity and recovery.
(3) Major Limitations
His-tag position, surrounding amino acid composition, and conformational exposure vary among proteins and can markedly affect binding efficiency to Co²⁺ media. Actual capacity therefore cannot be inferred from the metal type alone.
2.5 Metal-Chelating IDA and NTA Media
Metal-chelating IDA and NTA media contain pre-immobilized chelating groups but allow the experimenter to load the desired metal ions. These media are suitable for comparing different metal conditions such as Ni²⁺ and Co²⁺ or for developing specific IMAC methods. Compared with preloaded metal media, their use requires additional metal loading, washing, and regeneration steps, and consistency of metal loading directly affects batch-to-batch reproducibility.
Table 1 Major Differences Among Metal-Chelating Affinity Media
Medium System | Binding Characteristics | Major Advantages | Major Limitations | Typical Applications |
Ni-IDA | More open Ni²⁺ coordination sites | Relatively high binding capacity | Relatively pronounced nonspecific binding | Initial capture of highly expressed His-tagged proteins |
Ni-NTA | Balanced metal immobilization and His binding | Broad applicability | Relatively sensitive to chelators | Routine His-tag purification |
Co-NTA | Co²⁺-mediated high-selectivity binding | Lower background protein binding | Capacity generally lower than high-capacity Ni systems | High-purity His-tagged proteins |
Co Carboxymethyl Aspartate | Co²⁺ coordination-based | Expands screening options for Co systems | Large protein-to-protein differences in binding | Comparison of His-tag purification conditions |
Metal-Chelating IDA/NTA | User-loadable metal ions | Suitable for IMAC method development | Requires additional metal-loading steps | Screening of metal ions and ligands |
3 GST, MBP, and Strep-Tag Affinity Media
3.1 GST and Glutathione Affinity Media
(1) Binding Characteristics
GST fusion tags are captured through the affinity interaction between glutathione S-transferase and immobilized reduced glutathione and are subsequently eluted using free reduced glutathione as a competitive ligand.
(2) Major Advantages
GST systems provide relatively mild binding and elution conditions and can be used to preserve the native conformation and activity of target proteins. They can also be used directly in protein-interaction experiments such as GST Pull-down. GST fusion can additionally improve soluble expression of some recombinant proteins.
(3) Major Limitations
GST is a relatively large fusion tag and tends to form dimers, which may alter target-protein conformation, oligomeric state, or interaction characteristics. In structural studies and quantitative interaction assays, the GST tag often needs to be removed after purification.
3.2 MBP and Dextrin Affinity Media
(1) Binding Characteristics
MBP binds Dextrin- or Amylose-type polysaccharide ligands and can be eluted using competitive ligands such as Maltose. This system is mainly used for purification of MBP fusion proteins under native conditions.
(2) Major Advantages
MBP fusion can improve the solubility and folding state of some poorly soluble proteins and has substantial value for recombinant proteins that readily form inclusion bodies. Binding and elution are generally performed under relatively mild conditions.
(3) Major Limitations
MBP is a large fusion tag and may alter target-protein conformation, molecular weight, and complex formation. After MBP removal, free MBP, cleavage enzyme, and incompletely cleaved fusion protein also need to be further separated.
3.3 Strep-tag II Affinity Media
(1) Binding Characteristics
Strep-tag II is a short peptide tag that specifically binds Streptavidin-derived ligands such as Strep-Tactin. Twin Strep-tag increases binding strength by placing two tags in tandem.
(2) Major Advantages
Strep systems can bind under near-physiological conditions and can be gently eluted using competitive ligands such as Desthiobiotin. They are suitable for enzymes, protein complexes, and target proteins sensitive to high concentrations of imidazole or acidic conditions.
(3) Major Limitations
Biotin can occupy the binding sites of Strep-type ligands. High levels of free Biotin in samples can therefore reduce capture efficiency for tagged proteins. Samples containing Biotin culture supplements particularly require control of residual Biotin levels.
4 FLAG, HA, Myc, V5, and GFP Antibody Affinity Media
4.1 FLAG/DYKDDDDK, HA, Myc, and V5 Tag Media
(1) Binding Characteristics
FLAG/DYKDDDDK, HA, c-Myc, and V5 are epitope tags that can be captured using immobilized specific antibodies or nanobodies. Agarose media are suitable for column purification and batch incubation, whereas magnetic beads are suitable for IP, Co-IP, and low-volume sample processing.
(2) Major Advantages
Short peptide tags impose relatively little burden on target-protein molecular weight and structure, while antibody media provide high recognition specificity. Competitive elution using tag peptides can release target proteins and their interacting complexes under relatively mild conditions.
(3) Major Limitations
The capacity per unit volume of antibody media is generally lower than that of high-capacity IMAC media, and large-scale use is more expensive. Low-pH or denaturing elution may disrupt the activity of some proteins and may reduce the number of times the medium can be reused.
4.2 GFP Antibody and Nanobody Media
(1) Binding Characteristics
GFP fusion proteins can be captured using Anti-GFP antibodies or GFP Nanobodies, linking fluorescence imaging and expression detection with affinity enrichment.
(2) Major Advantages
GFP fusion constructs allow direct observation of expression and subcellular localization before purification, followed by IP, Co-IP, or affinity purification. Nanobodies are smaller molecules and can reduce some steric hindrance associated with conventional antibody media.
(3) Major Limitations
GFP is a relatively large fusion tag and may affect target-protein localization, conformation, or complex formation. Studies sensitive to the presence of the tag require validation using tag removal or alternative short-tag systems.
5 Boronic Acid Affinity Chromatography Media
5.1 Binding Mechanism
Phenylboronic acid-type ligands can form reversible boronate esters with 1,2-cis-diols or vicinal diol structures of other suitable configurations. Glycoproteins, carbohydrates, nucleosides, and some nucleotides can be enriched through this structural feature without prior introduction of an artificial protein tag.
5.2 Major Applications
Boronic acid affinity media are suitable for separating glycoproteins, glycan-containing biomacromolecules, and other compounds containing cis-diol structures. Glycoprotein capture depends on glycan structure rather than the primary amino acid sequence of the protein and can therefore be used for enrichment of glycoproteins from different sources and sample preparation for glycosylation-related analysis.
5.3 Influencing Factors
The ionization state of boronic acid groups and the efficiency of boronate ester formation are strongly affected by pH. Small molecules containing vicinal diol structures can compete for binding sites on the medium, while some proteins may also generate hydrophobic or electrostatic background. Changing pH or adding suitable polyhydroxylated competitor molecules can disrupt boronate ester formation and elute the target molecule.
6 Sulfated Affinity Media and Biomacromolecule Purification
6.1 Low-Sulfate Dextran Sulfate Media
(1) Binding Characteristics
Low-sulfate dextran sulfate media contain immobilized sulfated polysaccharides on their surfaces and exhibit heparin-mimetic molecular recognition and strongly negative charge characteristics. Target binding may include both affinity interactions with sulfated polysaccharides and ion-exchange interactions.
(2) Major Applications
These media are mainly used for separation of proteins and some plasma proteins capable of binding sulfated polysaccharides. Increasing salt concentration can weaken electrostatic interactions and thereby elute bound proteins.
(3) Major Limitations
Protein adsorption may arise from both specific affinity and nonspecific electrostatic interactions. In complex samples, salt gradients need to be used to observe separation between target and impurity peaks in order to determine whether the actual separation results from effective selectivity.
6.2 High-Sulfate Dextran Sulfate Media
(1) Binding Characteristics
High-sulfate dextran sulfate media contain a higher density of sulfate groups, which can enhance interactions with some viruses, virus-like particles, and other macromolecules with corresponding surface-binding characteristics.
(2) Major Applications
These media are mainly used for capture and enrichment of viruses and VLPs. Some viral surface proteins interact strongly with sulfated polysaccharides, allowing high-sulfate media to be incorporated into virus-purification processes.
(3) Major Limitations
Higher sulfate-group density can also increase electrostatic adsorption of other positively charged biomacromolecules. Because envelope composition, capsid structure, and surface proteins differ substantially among viruses, the binding capacity and selectivity of the same medium may vary considerably between different viruses.
6.3 Sulfate Ester-Modified Cellulose Affinity Media
(1) Binding Characteristics
Sulfate ester-modified cellulose media use sulfate ester groups on the cellulose matrix to generate a high-density negatively charged surface and capture biomacromolecules with corresponding binding characteristics through sulfated structures and electrostatic interactions.
(2) Major Applications
These media can be used to enrich certain viruses, biological particles, and macromolecular proteins. Compared with dextran sulfate agarose, the cellulose matrix has different pore structures, surface properties, and mass-transfer characteristics and can therefore produce different target-binding behavior.
(3) Major Limitations
High binding capacity may include both target-specific affinity interactions and general electrostatic adsorption. Salt concentration, pH, and target-particle size all affect actual separation performance.
Table 2 Major Differences Between Boronic Acid and Sulfated Affinity Media
Medium Type | Major Binding Basis | Typical Targets | Major Characteristics |
Boronic acid affinity | Cis-diol–boronate ester | Glycoproteins, carbohydrates, nucleosides | Enrichment using native vicinal diol structures |
Low-sulfate dextran sulfate | Heparin-mimetic interaction + electrostatic interaction | Proteins, some plasma proteins | Combines affinity and ion-exchange characteristics |
High-sulfate dextran sulfate | High-density sulfate-group interactions | Viruses, VLPs | Strong binding to some viral particles |
Sulfate ester-modified cellulose | Sulfated surface interactions | Viruses and other biomacromolecules | Cellulose matrix provides distinct mass-transfer characteristics |
7 Activated Affinity Media and Custom Ligand Coupling
7.1 CNBr-Activated Media
CNBr-activated media do not contain a predefined recognition ligand for a specific target. Instead, activated groups react with primary amino groups in proteins, antibodies, peptides, and other molecules. After ligand immobilization, recognition is determined by the coupled ligand itself, enabling construction of custom affinity systems based on antigen–antibody, receptor–ligand, protein–protein, and other interactions.
7.2 Ligand-Coupling Characteristics
CNBr coupling occurs mainly through primary amino groups in the ligand, so ligand orientation on the medium is generally random. If amino groups involved in coupling are close to the active region of the ligand, the binding site may become sterically masked. Increasing coupling density also does not necessarily continuously increase effective binding capacity because excessively high ligand density can create steric crowding and reduce accessibility for macromolecules.
7.3 4FF and Conventional Activated Media
4FF activated media use a mechanically stronger Fast Flow matrix and can be operated at higher flow rates and with larger column beds, making them more suitable for further scale-up. Conventional CNBr-activated media can be used for laboratory-scale ligand coupling, gravity columns, and batch affinity capture. In both cases, the final target-recognition characteristics are determined by the coupled ligand.
8 Medium Format and Purification Scale
8.1 Bulk Chromatography Media
Bulk agarose or other porous media can be used for self-packed columns, gravity columns, and low- to medium-pressure chromatography. Column-bed volume can be adjusted according to the amount of target protein, making these media suitable for method development, parameter comparison, and preparative protein purification.
8.2 Fast Flow Media
Fast Flow media increase mechanical strength through greater matrix crosslinking and can tolerate higher linear flow rates and column pressures. Excessively high flow rates reduce the diffusion and contact time of target molecules within the pores, so dynamic binding capacity may decrease as residence time becomes shorter.
8.3 Magnetic Affinity Media
Magnetic beads allow rapid capture, washing, and elution using a magnetic rack without column packing and are suitable for IP, Co-IP, low-volume protein enrichment, and parallel processing of multiple samples. Their major advantages are operational efficiency and recovery from small samples rather than large-volume preparative capacity.
8.4 Prepacked Columns
Prepacked columns have fixed bed heights and packing states and can reduce differences in flow distribution and column efficiency caused by manual packing. They are suitable for standardized workflows, repeated sample batches, and FPLC or low- to medium-pressure chromatography systems.
9 Binding, Washing, and Elution Conditions
9.1 Metal-Chelating Systems
His tags generally exhibit good metal-coordination ability under neutral to mildly alkaline conditions. Low concentrations of imidazole are used to suppress weakly binding host proteins, whereas high concentrations of imidazole competitively disrupt His-tag binding to Ni²⁺ or Co²⁺. Chelators such as EDTA may remove immobilized metal ions, and relatively high concentrations of reducing agents may also affect the stability of some IMAC media.
9.2 GST, MBP, and Strep Systems
GST is typically competitively eluted using reduced glutathione, MBP can be eluted using Maltose, and Strep systems can be eluted using competitive ligands such as Desthiobiotin. Competitive elution avoids strongly acidic or denaturing conditions, but residual elution ligands may affect subsequent enzymatic, binding, or quantitative assays.
9.3 Boronic Acid Affinity Systems
Boronic acid affinity binding depends on formation of reversible boronate esters between boronic acid groups and cis-diols. Reducing binding stability under appropriate conditions or adding polyhydroxylated compounds that compete for boronic acid binding sites can release captured molecules.
9.4 Sulfated Media
Electrostatic interactions in sulfated media can generally be weakened by increasing salt concentration. Salt gradients can be used to locate the target elution range while simultaneously assessing separation between nonspecifically bound components and the target peak.
9.5 CNBr-Coupled Media
Before use, CNBr-activated media require ligand coupling, blocking of unreacted active groups, and thorough washing. After coupling, the binding, washing, and elution systems are determined by the interaction between the immobilized ligand and target molecule.
10 Interpretation of Purification Results
10.1 High Levels of Target Protein in the Flow-Through Fraction
Substantial target protein in the Flow-through is generally associated with medium overload, insufficient tag exposure, absence of an effective affinity site in the target structure, or binding conditions that disrupt the interaction. When the medium is overloaded, increasing the column-bed volume can reduce flow-through loss. If the tag is buried by protein conformation, simply increasing the amount of medium will not restore binding.
10.2 Target Protein Loss in the Wash Fraction
Large amounts of target protein in the Wash fraction generally indicate that washing strength exceeds the binding stability of the target molecule. In His-tag systems, this is commonly caused by excessive imidazole concentration; in sulfated media, it may result from excessive salt concentration; and in antibody or other specific media, it may reflect a low-affinity ligand–target combination.
10.3 High Levels of Contaminants in the Elution Fraction
Background proteins in His-tag systems can be reduced by increasing wash imidazole concentration and ionic strength. Coelution from sulfated media is often associated with excessively broad electrostatic adsorption, whereas non-target molecules containing vicinal diol structures may compete with or co-bind the target in boronic acid media. Because contaminant sources differ, the corresponding mechanisms for impurity removal also differ.
10.4 High Purity but Low Recovery
High purity accompanied by low recovery is commonly associated with insufficient binding during sample loading, overly stringent washing, or incomplete elution. Comparing target-protein distribution among the Input, Flow-through, Wash, and Elution fractions can identify the stage at which target loss occurs.
10.5 Reduced Protein Activity After Elution
High concentrations of imidazole, acidic elution, high salt, competitive ligands, and prolonged purification may alter the conformation or complex state of some proteins. When enzyme activity decreases despite normal SDS-PAGE purity, elution conditions and the protein buffer environment need to be further examined rather than evaluating purification solely by purity.
11 Affinity Chromatography Media and Supporting Products
11.1 His-Tag and Metal-Chelating Affinity Media
Catalog # | Product Name | Grade & Purity | Main Application |
Ni-IDA Affinity Chromatography Medium 6FF (His Tag) | BioReagent, 50% v/v; 45-165 μm | High-capacity purification of His-tagged proteins | |
IMAC Agrose (NTA) 6FF | BioReagent, 70% v/v; 90μm | Construction of customized metal-ion NTA affinity systems | |
Chelating Agrose (IDA) 6FF | BioReagent, 70% v/v; 90 μm | Construction of customized metal-ion IDA affinity systems | |
Co-NTA Affinity Chromatography Medium 6FF (His Tag) | BioReagent, 50% v/v | High-selectivity purification of His-tagged proteins | |
Cobalt CM-Asp Affinity Resin (His tag) | BioReagent, 50% v/v; 30-100 μm | Co²⁺-based affinity purification of His-tagged proteins | |
Ni-NTA Affinity Resin (His tag) | BioReagent, for protein analysis, 30-35 mg His-Tagged protein/mL wet gel; 75 μm | Routine affinity purification of His-tagged proteins | |
UltraBio™ His-tag Purification Resin |
| His-tagged protein purification under specialized buffer conditions | |
UltraBio™ TED-Ni Magnetic Agarose Beads | See COA | Microscale and high-throughput purification of His-tagged proteins | |
His-Tag Protein Purification Kit (Ni-IDA, pH 8.0) | BioReagent, for protein analysis | Standardized Ni-IDA purification | |
His Pull-Down Kit (Agarose beads) | BioReagent, for IP, ready-to-use | His-tag pull-down and enrichment of interacting proteins | |
Ni-Agarose Column In-Place Cleaning Kit | BioReagent, Sterile-filtered | Cleaning and maintenance of Ni-affinity media |
11.2 GST-, MBP-, and Strep-Tag Affinity Products
Catalog # | Product Name | Grade & Purity | Main Application |
Glutathione Agarose Resin (GST tag) | BioReagent, 50% v/v | Affinity purification of GST fusion proteins | |
GST Pull-Down Kit (Magarose Beads) | BioReagent | GST pull-down assays | |
Dextrin Agarose Resin (MBP tag) | BioReagent, endotoxin tested, 50% v/v | Purification of MBP-tagged fusion proteins | |
MBP-Tag Protein Purification Kit (pH 7.4) | BioReagent, for Protein analysis | Standardized purification of MBP-tagged proteins | |
MBP purification system (pH 8.0) | BioReagent, for Protein analysis | Standardized purification of MBP-tagged proteins | |
UltraBio™ Anti-MBP Agarose | BioReagent, for IP, 50% v/v | IP, Co-IP, and purification of MBP-tagged proteins | |
Strep-tag II Agarose Purification Resin 4FF | BioReagent, 50% v/v | Purification of Strep-tag II and Twin Strep-tag proteins | |
UltraBio™ Strep Tactin Magnetic Agarose Beads | BioReagent, 20% v/v; particle size: 10-37 μm | Magnetic purification of Strep-tagged proteins | |
Streptavidin Beads 6FF | BioReagent, 50% v/v | Affinity capture of biotinylated molecules |
11.3 Boronate and Sulfated Affinity Media
Catalog # | Product Name | Grade & Purity | Main Application |
Boronate Affinity Chromatography Resin | BioReagent, 50% v/v; 30~100 μm | Enrichment of glycoproteins and cis-diol-containing molecules | |
Dextran Sulfate Affinity Resin (Low Sulfur / For Protein Purification) | BioReagent, 50% v/v; 30~100 μm | Affinity separation of proteins and plasma proteins | |
Dextran Sulfate Affinity Resin (High Sulfur / For Virus Purification) | BioReagent, 50% v/v; 30~100 μm | Capture and purification of viruses and VLPs | |
Sulfate-modified Cellulose Affinity Resin | BioReagent, 50% v/v; 45~165 μm | Purification of viruses and other biomacromolecules |
11.4 Activated Affinity Chromatography Media
Catalog # | Product Name | Grade & Purity | Main Application |
CNBr-activated Agrose 4FF | BioReagent, 70% v/v; 90μm | Fast Flow medium for customized ligand coupling | |
CNBr-activated Agrose | BioReagent, 50% v/v; 90μm | Coupling of antibodies, proteins, and peptide ligands |
11.5 FLAG-, HA-, Myc-, V5-, and GFP-Tag Products
Catalog # | Product Name | Grade & Purity | Main Application |
Anti-HA Agarose Resin | BioReagent, 50% v/v | HA-tagged protein purification and IP | |
UltraBio™ Anti-HA Magnetic Beads | BioReagent, for IP, 10 mg/mL, Particle Size: 1 μm | IP and Co-IP of HA-tagged proteins | |
Anti-Myc Affinity Gel | BioReagent, 50% v/v | c-Myc-tagged protein purification and IP | |
UltraBio™ Anti-Myc Magnetic Beads | BioReagent, for Protein analysis, 10 mg/mL; Particle Size: 1 μm | Magnetic IP and purification of Myc-tagged proteins | |
UltraBio™ Anti-DYKDDDDK Agarose | BioReagent, for IP, 50% v/v | Purification of FLAG/DYKDDDDK-tagged proteins | |
UltraBio™ Anti-DYKDDDDK tag (Binds to Flag tag) Magnetic Beads | BioReagent, for IP, 10 mg/mL, Particle Size: 1 μm | Magnetic capture of FLAG-tagged proteins | |
Flag-tag Protein IP Assay Kit (Agarose Gel) | BioReagent, Ready-to-use, for IP | FLAG-tagged protein IP and Co-IP | |
Flag-tag Protein IP Assay Kit (Nanobody Magnetic Beads Method) | BioReagent, Ready-to-use, for IP | FLAG Nanobody magnetic-bead IP and Co-IP | |
Anti-GFP Agarose Resin | BioReagent, 50% v/v | Purification and IP of GFP/EGFP-tagged proteins | |
Anti-GFP Nanobody Magnetic Beads | BioReagent, Magnetic Bead Content: ≥10% (V/V); Bead Diameter: 10-30 μm | Magnetic affinity capture of GFP-tagged proteins | |
UltraBio™ Anti-V5 Magnetic Beads |
| IP, Co-IP, and purification of V5-tagged proteins | |
V5-tag IP/Co-IP Kit (Nanobody Magnetic Beads) | BioReagent, Ready-to-use, for IP | IP and Co-IP of V5-tagged proteins |
Affinity chromatography media mainly include metal-chelating, tag-ligand, antibody-affinity, boronic acid-affinity, sulfated-affinity, and activated coupling systems. Ni²⁺/Co²⁺ media are mainly used for His-tagged proteins; glutathione, Dextrin, and Strep-type media are used for GST, MBP, and Strep tags, respectively; boronic acid media use cis-diol structures for enrichment; sulfated media are used for capture of certain proteins and viral particles; and CNBr-activated media are used to construct custom affinity systems.
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