Validation Data Gallery

  • Model fig of Spot peptide.

    Model fig of Spot peptide.

  • Purification of Spot-tagged GFP with Spot-Cap from HEK293T cell lysate in gravity flow column format. Elution 1-6: 6 fractions à 2 CVs Spot-peptide (100 μM) in PBS.

    Purification of Spot-tagged GFP with Spot-Cap from HEK293T cell lysate in gravity flow column format. Elution 1-6: 6 fractions à 2 CVs Spot-peptide (100 μM) in PBS.

  • Workflow of Spot-Cap purification.

    Workflow of Spot-Cap purification.

  • Spot-Cap has a higher selectivity than anti-DYKDDDDK resin. While Spot-Cap is optimized for minimal contamination of host cell proteins, the anti-DYKDDDDK resin binds significant amounts of mammalian proteins. Incubation of control HEK293T cell lysate expressing no tagged protein with Spot-Cap or anti-DYKDDDDK resin. Elution with 100 μM Spot-peptide or 0.1 g/L DYKDDDDK-peptide. Similar results were obtained for yeast, bacteria, and insect cell lysates (data not shown).

    Spot-Cap has a higher selectivity than anti-DYKDDDDK resin. While Spot-Cap is optimized for minimal contamination of host cell proteins, the anti-DYKDDDDK resin binds significant amounts of mammalian proteins. Incubation of control HEK293T cell lysate expressing no tagged protein with Spot-Cap or anti-DYKDDDDK resin. Elution with 100 μM Spot-peptide or 0.1 g/L DYKDDDDK-peptide. Similar results were obtained for yeast, bacteria, and insect cell lysates (data not shown).

  • The elution efficiency of Spot-Cap is constant even after 4 regeneration cycles. Elution of Spot-tagged PCNA with Spot-peptide. R0: Reference (elution fraction of Spot-PCNA without regeneration); R1-R4: Elution fraction of Spot-PCNA after 1, 2, 3, and 4 regeneration cycles with 100 mM glycine pH 2.0.

    The elution efficiency of Spot-Cap is constant even after 4 regeneration cycles. Elution of Spot-tagged PCNA with Spot-peptide. R0: Reference (elution fraction of Spot-PCNA without regeneration); R1-R4: Elution fraction of Spot-PCNA after 1, 2, 3, and 4 regeneration cycles with 100 mM glycine pH 2.0.

  • Purification of Spot-tagged GFP with Spot-Cap from HEK293T cell lysate in batch format. Elution: 6 fractions à 2 BVs Spot-peptide (100 μM) in PBS.

    Purification of Spot-tagged GFP with Spot-Cap from HEK293T cell lysate in batch format. Elution: 6 fractions à 2 BVs Spot-peptide (100 μM) in PBS.

  • Application of Spot-tag related products.

    Application of Spot-tag related products.

ChromoTek Spot-peptide

Spot-peptide
Cat No. ep

Peptide

Spot-Tag®

Applications

Elution

Conjugate

Unconjugated

Spot peptide

Size/Concentration: 

-/ -


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Product Information

Spot-peptide

ApplicationsElution of Spot-tagged fusion proteins bound to Spot-Cap
SequencePeptide of amino acid sequence PDRVRAVSHWSS
FormLyophilized powder
ConjugateUnconjugated
Molecular weight1,397 g/mol
Purity>96% (HPLC)
ReconstitutionDissolve Spot-peptide in PBS (1 mg Spot-peptide in 140 µL PBS,
10 mg Spot-Peptide in 1.4 mL PBS) to a final concentration of
5 mM (7 mg/mL). Aliquot upon reconstitution.
DilutionDilute Spot-peptide to 0.1 mM in PBS for elution of Spot-tagged
proteins from Spot-Cap.
Storage ConditionUpon receipt store at -20°C/-4°F.
Aliquot upon reconstitution and store at -20°C/-4°F.
Avoid repeated freeze / thaw cycles.
ShipmentShipped at ambient temperature. Upon receipt store at -20°C.
Aliquot upon reconstitution and store at -20°C. Avoid repeated
freeze / thaw cycles.
Spot-Tag® OriginEngineered 12 amino acid sequence modified from the unstructured N-terminus of beta-catenin
Cross ReactivityThe Spot-Nanobody shows minimal cross reactivity with beta-catenin.

Publications

ApplicationTitle

Cells

Cep192, a Novel Missing Link between the Centrosomal Core and Corona in Dictyostelium Amoebae.

Authors - Valentin Pitzen