Affinity optimized short peptide tag

Spot-Tag & Applications

Affinity Optimized Short Peptide Tag

The Spot-Tag^® has the affinity-optimized 12 amino acid sequence PDRVRAVSHWSS. Spot-Nanobodies bind strongly to the Spot-Tag. The inert tag sequence can be added to the N-terminal and to the C-terminal side of the protein of interest. This works by PCR using modified primers or by using any of the Spot vectors offered (click here for more information). The expression of Spot-tagged proteins has been successfully tested in bacteria, yeast, mammalian cell lines, plant and insect cells.

Spot-Nanobody advantages compared to other antibodies:	Spot-Tag advantages compared to other peptide tags:
More robust & stable than IgG antibodies High affinity of 6 nM to Spot-tagged proteins and 0.7 nM to Spot-Tag Less background than other tested affinity resins Less unspecific binding than other tested capture and detection antibodies Higher resolution than conventional antibodies Better epitope accessibility and tissue penetration than conventional IgG antibodies Effective and gentle elution at 4°C (Spot-Cap)	Less negatively charged than FLAG^®-, Myc-, and HA-tags Short Inert, unstructured tag does not interference with fusion protein structure and function Binding of a single tag Suitable for ubiquitination assays: comprises no lysine

Spot-Nanobody advantages
compared to other antibodies:

Spot-Tag advantages
compared to other peptide tags:

More robust & stable than IgG antibodies
High affinity of 6 nM to Spot-tagged proteins and 0.7 nM to Spot-Tag
Less background than other tested affinity resins
Less unspecific binding than other tested capture and detection antibodies
Higher resolution than conventional antibodies
Better epitope accessibility and tissue penetration than conventional IgG antibodies
Effective and gentle elution at 4°C (Spot-Cap)

Less negatively charged than
FLAG^®-, Myc-, and HA-tags
Short
Inert, unstructured tag does not interference with fusion protein structure and function
Binding of a single tag
Suitable for ubiquitination assays: comprises no lysine

Overview of Spot related products by application

Immunoprecipitation (IP): use Spot-Trap

Spot-Trap^® is the Spot-Nanobody coupled to agarose or magnetic agarose beads.

NO contaminating heavy & light antibody chains
Harsh washing conditions
Native & non-native elution
High affinity, stringent wash

Protein purification: use Spot-Cap

Spot-Cap^® is an optimized Spot-Nanobody derivative coupled to agarose beads for purification of Spot-tagged proteins.

One step protein purification
High binding capacity
Multiple regeneration cycles
Effective elution at 4°C with Spot-peptide or pH shift

Immunofluorescence (IF) & Western blotting (WB): use Spot-Label

Spot-Label^® is the Spot-Nanobody conjugated to fluorescent dyes. The bivalent Spot-Label is the first Nanobody/ peptide-tag that was applied in super-resolution microscopy.

Higher resolution
Better epitope access & tissue penetration
Very small probe size

Expression of Spot-tagged fusion proteins: use Spot-Tag vectors

A selection of Spot-Tag vectors is available. Alternatively, Spot-Tag can be easily cloned by PCR primers.

Elution of bound Spot-tagged protein: use Spot peptide

Spot peptide for gentle elution of Spot-tagged proteins from Spot-Cap™.

Western Blotting, ELISA: use Spot VHH, recombinant binding protein

Non-conjugated Spot-Nanobody is called Spot VHH, recombinant binding protein and can be used for:

Very sensitive in Western blot and ELISA
Coupling of dyes, biotin, etc. via NHS Ester reaction

For more information about the Spot-Tag and Spot Nanobody see

Spot Capture and Detection System
• About Spot
• Spot Nanobody

Free samples available

Request free sample here and test Spot in your experiment.