V5 Epitope‑Tag: Versatile Peptide Labeling Toolkit for Modern Molecular Biology Research
Molecular Origin and Core Biochemical Traits of the V5 Epitope‑Tag
The V5 epitope‑tag originates from a 14‑residue peptide segment (GKPIPNPLLGLDST) within SV40 small‑T antigen P‑kappa mutant polypeptide sequences. This short peptide reaches approximately 1.5 kDa molecular mass, featuring evenly distributed hydrophilic amino acid residues to limit non‑specific intermolecular interactions. Its linear epitope architecture grants antibody‑binding capacity independent of disulfide‑bond formation or intact tertiary protein folding states. Researchers can genetically fuse this peptide tag to either N‑terminal or C‑terminal positions of target recombinant proteins according to experimental design requirements. N‑terminal fusion frequently pairs with signal peptide sequences for secreted recombinant protein research, while C‑terminal tagging suits most intracellular protein characterization workflows. Owing to its compact dimensions, V5 tag imposes minimal steric hindrance and seldom disturbs native folding, ligand‑binding capacity or physiological activity of fusion partner proteins. This property renders it broadly compatible with yeast, mammalian HEK293‑CHO, and baculovirus‑driven insect‑cell expression platforms.
Distinct Experimental Advantages Compared With Other Widely‑Used Epitope Tags
Unlike large fluorescent fusion partners such as GFP, V5 peptide avoids intrinsic fluorescence interference during cellular imaging and functional readout acquisition. The linear epitope maintains stable antibody‑recognition performance under denaturing SDS‑PAGE sample conditions required for standard Western blot workflows. It can also operate under native buffer environments for immunoprecipitation and protein‑complex enrichment experiments. V5 reagents frequently combine with His‑tag or FLAG‑tag sequences to construct dual‑tag recombinant constructs satisfying multi‑step detection and purification workflows. Anti‑V5 antibodies exhibit low non‑specific background against endogenous cellular proteomes, lowering false‑positive signal occurrences within complex cell‑lysate matrices.
Multi‑Modal Detection Workflows Enabled by V5‑Tagging Systems
Western‑blot immuno‑detection with anti‑V5 antibodies enables sensitive measurement of target protein abundance even for low‑abundance recombinant protein specimens. Conjugated HRP or FITC anti‑V5 variants streamline downstream signal development without secondary‑antibody incubation steps. Immunoprecipitation and co‑immunoprecipitation constitute another major application field for capturing V5‑labelled bait proteins and their physiological interaction partners. Mild elution schemes using competitive V5‑peptide or low‑pH glycine buffer preserve assembled protein‑complex integrity for subsequent LC‑MS interactome profiling. Immunofluorescence microscopy permits precise visualization of V5‑tagged protein subcellular localization across cytoplasmic, membrane and nuclear cellular compartments. When paired with flow cytometry, V5‑tag detection quantifies surface‑expressed recombinant protein abundance within mixed transfected‑cell populations.
Affinity Purification Advantages Using V5‑Immunoaffinity Bead Platforms
V5‑based immunoaffinity purification employs antibody‑coupled agarose or magnetic beads to isolate V5‑fusion proteins from crude cell lysate or culture supernatant material. This capture mechanism does not depend on transition‑metal ions required for conventional His‑tag nickel‑chelating purification protocols. Removing metal‑ion exposure circumvents metal‑triggered oxidative damage and aberrant aggregation for redox‑sensitive recombinant protein substrates. Operators can select from multiple gentle elution alternatives including competitive peptide displacement, low‑pH buffer elution or mild‑detergent containing buffer systems. Magnetic bead formats simplify handling for small‑scale batch purification while agarose bead variants support larger‑volume preparative recombinant‑protein isolation runs. This toolkit suits membrane‑protein, transcription‑factor and multi‑subunit protein‑complex sample preparation for crystallography or cryo‑EM sample‑preparation pipelines.
Expanded Applications Within Functional Genomics and CRISPR‑Assisted Research
High‑throughput ORFeome library projects deploy genome‑wide V5‑tagged open‑reading‑frame collections for large‑scale protein‑localization and interactome screening assays. V5‑tagging combines with CRISPR‑Cas9‑mediated homology‑directed repair to install epitope‑tags onto endogenous genomic loci. Endogenous tagging enables protein measurement under native physiological promoter‑driven expression levels, eliminating artifacts originating from strong plasmid‑based overexpression systems. V5‑labelled bait constructs also serve in proximity‑labelling experimental workflows such as APEX2‑V5 fusion setups for mapping spatially‑restricted subcellular proteome landscapes. RNA‑interference rescue assays utilizing V5‑tagged rescue constructs assist researchers in verifying gene‑knockdown phenotype specificity and excluding potential off‑target‑effect confounders.
Forward‑Looking Technical Development Trends for V5‑Tagging Toolkits
Ongoing technical innovations advance V5‑tag systems toward higher‑sensitivity detection and multi‑modal experimental compatibility. Nanobody‑derived anti‑V5 binders deliver improved tissue penetration kinetics for fixed‑sample high‑resolution imaging research. Environment‑responsive V5‑tag variants are under evaluation to report intracellular pH and redox‑state fluctuations within living‑cell experimental models. Combined with microfluidic single‑cell‑analysis platforms, V5‑immuno‑detection achieves multiplex protein quantification at single‑cell resolution. Synthetic‑biology workflows treat V5‑tag sequences as standardized BioBrick components to streamline modular genetic‑circuit assembly and validation work.
Comprehensive V5‑Tag Antibody and Affinity Bead Reagents from ANT BIO PTE. LTD
ANT BIO PTE. LTD supplies a complete portfolio of unconjugated, fluorophore‑ or enzyme‑conjugated anti‑V5 antibodies alongside immunoaffinity agarose and magnetic‑agarose beads. Every reagent undergoes multi‑assay validation across WB, IF, flow cytometry and immunoprecipitation workflows to guarantee stable experimental performance.
Catalog Table of V5‑Tag Detection & Purification Reagents
| Catalog Number | Full Product Name | Core Product Specifications | Available Pack Sizes |
|---|---|---|---|
| S0B0202 | V5 Tag Mouse mAb (FITC Conjugate) (S‑610‑66) | FITC‑labelled mouse monoclonal anti‑V5‑tag | 25 μL / 100 μL / 1 mL |
| S0B1737 | Rabbit Anti‑V5 tag agarose Beads | Antibody‑coupled agarose for V5‑fusion protein affinity enrichment | 125 μL / 250 μL / 500 μL / 1 mL / 5 mL / 10 mL |
| S0B1720 | Rabbit Anti‑V5 Tag Magnetic Agarose | Magnetic‑bead‑based anti‑V5 affinity resin | 125 μL / 250 μL / 500 μL / 1 mL / 5 mL / 10 mL |
| S0B0195 | V5 Tag Mouse mAb (HRP Conjugate) (S‑610‑66) | HRP‑conjugated mouse monoclonal anti‑V5‑tag | 25 μL / 100 μL / 1 mL |
| S0B0537 | V5 Tag Recombinant Rabbit mAb (S‑609‑13) | Unconjugated recombinant rabbit monoclonal anti‑V5‑tag | 25 μL / 100 μL / 1 mL |
| S0A0140 | Schistosoma japonicum GST Protein, V5 tag | E.‑coli‑expressed V5‑GST fusion control antigen | 10 μg / 25 μg / 50 μg |
Functional Validation Characteristics of ANT BIO PTE. LTD V5‑Tag Reagents
Peptide‑array screening confirms minimal cross‑reactivity against endogenous mammalian proteome epitopes to reduce experimental background noise. Recombinant antibody clones exhibit consistent batch‑to‑batch performance compared to traditional hybridoma‑derived antibody lots. Conjugated HRP and FITC formats eliminate secondary‑antibody incubation steps and shorten hands‑on experimental processing duration. Both agarose and magnetic affinity‑bead variants retain high V5‑tag capture capacity under native and mild‑denaturing lysis‑buffer environments. The S0A0140 V5‑GST fusion protein serves as convenient positive control material for antibody performance qualification assays.
Core Fundamental Research Applications for V5‑Tag Reagent Panel
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Western‑blot quantification for assessing V5‑fusion recombinant‑protein expression levels in transient or stable transfected cell lines
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Immunoprecipitation and co‑IP workflows for capturing V5‑bait protein complexes for subsequent interactome LC‑MS/MS profiling
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Immunofluorescence cellular imaging to characterize subcellular distribution patterns of transiently or endogenously V5‑tagged target polypeptides
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Magnetic‑bead or agarose‑based affinity purification for preparative isolation of V5‑fusion proteins for structural‑biology sample‑preparation
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Flow‑cytometry quantitative detection for surface‑localized V5‑tagged transmembrane receptor proteins within mixed cell populations
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CRISPR‑HDR endogenous tagging experimental validation for monitoring native‑level protein abundance and localization dynamics
Global Manufacturing & Compliance Standards
All V5‑tag antibody and bead reagents complete multi‑platform functional validation before commercial release. Manufacturing facilities maintain ISO9001, ISO13485 and EU 98/79/EC certification standards for life‑science‑research‑reagent production. In‑house application science teams supply optimized immunodetection and affinity‑purification SOP documents alongside curated reference publications. The broader reagent ecosystem integrates PTM‑modification antibodies, ELISA kits and cell‑separation magnetic beads for unified multi‑omics molecular‑biology pipelines.
ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs
At ANT BIO PTE. LTD., we are committed to advancing life science research through high‑quality, reliable reagents and comprehensive solutions. Our specialized sub‑brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer‑centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.
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