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An Updated Overview of Protein Tags

L L • Apr 24, 2019
A helpful guide for the variety of different protein tags - trending in recent growth

An Updated Overview of Protein Tags

A guide to recent and growing trends in protein tags


Introduction

Protein tagging has been around for decades, but their traditional uses are not frequent enough to make them common lab knowledge. Recent improvements in biochemical techniques have been increasing their frequency as creative design and analysis tools. In this overview, we offer a guide for the variety of different protein tags that are trending in recent growth.

This guide will cover:

  • Principle Affinity Tags
  • Biotin Tags
  • Epitope Tags
  • Fluorescent Tags


Principle Affinity Tags

Arg-tag

Overview
Function Immobilize functional proteins on flat surface
Application Studying interactions with ligands
Feature GFP with Arg-tag can be bound to mica surfaces
Application Scanning probe microscopy applications
Terminus Attachment C-term
Compatible Domain Bacterial
Measured Biochemistry
Purity 95%
Yield 44%


His-tag

Overview
Function Immobilization, linear recognition motif
Application Purification (most common method)
Feature Strongest interaction with immobilized metal ion matrice
Application Immobilized metal-affinity chromatography (IMAC)
Terminus Attachment N or C-terminal
Compatible Domain Bacterial; Yeast; Mammalian; Viral
Measured Biochemistry
Purity 95%
Yield 44%


GST

Overview
Function Increased solubility, highly antigenic
Application Purification
Feature Amplified detection
Application Protein expression
Terminus Attachment N or C-terminal


Biotin Tags


Strep-tag

Overview
Function Matrix regenerable, recognition motif
Application Purification and detection
Feature Eukaryotic cell surface display, surface immobilization
Application SPR CHIPS, NMR, Crystallization, Protein interactions
Terminus Attachment N or C-terminal
Compatible Domain Bacterial; Yeast; Mammalian; Insects; Plants; Synthetic


Streptavadin-tag

Overview
Function Increase proteolytic stability, high affinity immobilization
Application Detection and Expression support
Feature Newer low affinity versions
Application Purification
Terminus Attachment N or C-terminal
Compatible Domain Bacterial; Yeast; Mammalian; Insects; Plants; Synthetic


Epitope Tags


c-myc-tag

Overview
Function 9E10 immunological recognition
Application Immunoblotting/WB, Immunoprecipitation, Flow Cytometry
Feature Convertible Reagent
Application Protein engineering
Terminus Attachment N or C-terminal
Compatible Domain Bacterial; Yeast; Mammalian; Insects


FLAG-tag

Overview
Function Peptide bound to antibody (M1)
Application Purification
Feature Consistent non-denaturing conditions
Application Purifying active fusion proteins
Terminus Attachment N or C-terminal
Compatible Domain Bacterial; Yeast; Mammalian
Measured Biochemistry
Purity 90%


HA-tag

Overview
Function Viral antibody recognition
Application Detection
Feature Anti-HA antibodies specific
Application Expression
Terminus Attachment N or C-terminal
Compatible Domain Mammalian
Measured Biochemistry
Yield Low


Fluorescent Tags


Luciferase

Overview
Function Luminescent dye
Application Detection
Feature Serve as a reporter immediately upon translation
Application In-situ hybridization, RNA processing, RNA transfection
Terminus Attachment N-terminal
Compatible Domain Bacterial; Yeast; Mammalian; Insects


GFP

Overview
Function Intrinsic fluorescence can permit native detection
Application Detection
Feature No antibody needed for detection
Application Monitor gene expression, protein folding, and targeting
Terminus Attachment N-terminal
Compatible Domain Bacterial; Yeast; Mammalian; Insects


S-tag

Overview
Function Specific linear recognition motif
Application Detection
Feature RNase S specificity
Application Diverse assays; quantitative, colorimetric no antibody
Terminus Attachment N or C-terminal
Compatible Domain Bacterial; Yeast; Mammalian


Points to Consider

  1. FLAG mAb purification systems have not always been stable and often require expert intervention.
  2. The presence of C-terminal tags can potentially contribute to a protein's loss of function. Particularly in the case of enzymes.
  3. Streptavadin-binding peptide (SBP) is only attachable to the C-terminus
  4. S-tag in low pH elution may alter protein properties and limit the matrix reproducibility
  5. anti-myc antibody antibody purification is inconsistent and has been proven to result in poor yields


References

  1. Nilsson et al. (1997a)
  2. Lazzaroni et al. (1985)
  3. Goldstein et al. (1992)
  4. Crespo et al. (1997)
  5. Makrides (1996)
  6. McLean et al. (2001)
  7. Nilsson et al. (1997b)
  8. Wang et al. (1996)
  9. Podbielski et al. (1992)
  10. Jones et al. (1995)
  11. Morandi et al. (1984)
  12. Kaldalu et al. (2000)
  13. Jones et al. (1995)
  14. Rubinfeld et al. (1991)
  15. Smith (2000)
  16. Gerdes and Kaether (1996)
  17. Los et al. (2008)
  18. Terpe (2003)
  19. Imagawa et al. (1982)
  20. Fritze and Anderson (2000)
  21. Tai et al. (1988)
  22. Kuliopulos and Walsh (1994)
  23. Kwatra et al. (1995)
  24. Tai et al. (1988)
  25. Karp and Oker-Blom (1999)
  26. Kolodziej and Young (1991)
  27. Terpe (2003)
  28. Kimple and Sondek (2002)
  29. Stevens (2000)
  30. Bornhorst and Falke (2000)
  31. Stevens (2000)
  32. Abdulaev et al. (2005), Biao et al. (2004), Ruan et al. (2004)
  33. Fritze and Anderson (2000)
  34. Berlot (1999)
  35. Nilsson et al. (1997b)
  36. Skerra and Schmidt (2000)
  37. Sano et al. (1998)
  38. Terpe (2003)
  39. Stevens (2000)
  40. Li (2011)
  41. Terpe (2003)
  42. Nelson et al. (1999)
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