Selection guide: peptide tags
Do you know which tag is best for your protein? Explore the options below to choose the best tag for your protein and application.
His tag The most versatile tag for protein purification |
Express pET Express & Purify Kits »
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Purify His60 nickel resin » TALON cobalt resins »
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Detect 6xHis Monoclonal Antibody » 6xHN Polyclonal Antibody »
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Features
- Distinct types of his tags are available, including the 6xHN tag (12 amino acids)
- Most common affinity tag used to purify proteins
- Binds to coordinated metals such as Ni2+ or Co2+
- His-tagged proteins can be eluted with imidazole or low-pH buffer
- A his tag can be incorporated on either the N- or C-terminus
- Can be expressed in bacterial, yeast, mammalian, and baculovirus-infected insect cells or in vitro
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When to Use
- When a small tag is required to minimize effects on protein function
- When a low metabolic load is needed to maintain normal cell physiology and/or to increase protein expression
- When a high-capacity affinity resin is needed to increase yields and reduce costs
- To reduce the cost of affinity purification by using an inexpensive resin
- To reduce the cost of affinity purification by using a resin that can be regenerated multiple times
- When the choice to purify a protein under native or denaturing conditions is needed
- When mild elution conditions are necessary
- For large-scale and HTP purifications
- To study protein-protein or nucleic acid-protein interactions involving an immobilized his-tagged protein
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When Not to Use
- When the tagged protein sample is in the presence of other metals or chelating reagents (EDTA)
- When eluting proteins with imidazole buffers will interfere with downstream processes such as crystallography or NMR applications (elution with low pH buffers or desalting may overcome this issue)
- When high concentrations of reducing agents (DTT) are necessary
- When trying to purify metalloproteins
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Myc tag A small, immunoreactive tag—ideal for co-IP & western blots |
Express pCMV-Myc & pCMV-HA Vector Set »
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Detect c-Myc Monoclonal Antibody »
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Features
- Small peptide tag (11 amino acids)
- Binds to the c-Myc Monoclonal Antibody
- Myc-tagged protein can be eluted with low pH buffer or sample buffer
- A Myc-tag can be incorporated on either the N-terminus, the C-terminus, or internally
- Can be expressed in bacterial, yeast, mammalian, and baculovirus-infected insect cells
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When to Use
- When a small tag is required to minimize effects on protein function
- When a low metabolic load is needed to maintain normal cell physiology and/or to increase protein expression
- To monitor recombinant protein expression in bacterial, yeast, mammalian, or insect cells
- For co-immunoprecipitation studies, Western blots, and flow cytometry
- To monitor subcellular localization of the tagged protein in mammalian cells
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When Not to Use
- When an inexpensive purification resin is needed
- When low pH elution conditions can damage the target protein or affect its functionality
- For applications requiring a resin with a high binding capacity or high yields
- When matrix stability/shelf-life is a factor
- For large-scale and HTP purifications
- When regeneration of the matrix is desired
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HA tag Ideal for co-IP and western blots—unlikely to interfere with the function of your protein |
Express pCMV-Myc & pCMV-HA Vector Set »
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Features
- Small peptide tag (9 amino acids)
- Binds to the HA Tag Polyclonal Antibody
- An HA tag can be incorporated on either the N-terminus, the C-terminus, or internally
- Can be expressed in bacterial, yeast, mammalian, and baculovirus-infected insect cells
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When to Use
- When a small tag is required to minimize effects on protein function
- When a low metabolic load is needed to maintain normal cell physiology and/or to increase protein expression
- To monitor recombinant protein expression in bacterial, yeast, mammalian, or insect cells
- For co-immunoprecipitation studies and western blots
- To monitor subcellular localization of the tagged protein in mammalian cells
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When Not to Use
- As an affinity tag to purify tagged protein
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GST tag Binds with high affinity and specificity |
Purify GST-tag resins »
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Features
- Large protein tag (glutathione S-transferase, 26 kDa protein)
- Binds to glutathione
- GST-tagged proteins can be eluted with reduced glutathione
- A GST tag can be incorporated on either the N- or C-terminus
- Can be expressed in bacterial, yeast, mammalian, and baculovirus-infected insect cells
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When to Use
- To reduce the cost of affinity purification by using an inexpensive resin
- To reduce the cost of affinity purification by using a resin that can be regenerated multiple times
- When mild elution conditions are necessary
- To protect some recombinant proteins from intracellular protease cleavage
- When it is necessary to use a tag that may enhance the solubility of the tagged protein
- When removal of the tag is necessary
- To study protein-protein or protein-nucleic acid interactions
- To use as an antigen for immunology or vaccination studies
- When the protein needs to be purified under strong reducing conditions
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When Not to Use
- When a low metabolic load is needed to maintain normal cell physiology and/or to increase protein expression
- When a small tag is required
- When a homodimeric protein tag is not suitable
- When trying to isolate an oligomeric protein
- When purification under denaturing conditions is required
- When the target protein contains disulfides which may be reduced by the elution buffer
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