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ChromoTek Spot-Cap™ and Peptide

Spot-Cap® is a Nanobody-based affinity resin optimized for one-step protein purification of Spot-tagged proteins.

Specificity

Spot-Tag

Applications

Protein purification

Conjugate

Unconjugated

Cat no : eca-ep

Synonyms

Spot Affinity Gel; Spot Affinity Matrix

Validation Data Gallery

  • 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.

Product Information

Spot-Cap® is a Nanobody-based affinity resin optimized for one-step protein purification of Spot-tagged proteins.

ApplicationsProtein purification
ClonalityMonoclonal
HostAlpaca
Specificity/TargetSpot-Tag PDRVRAVSHWSS
Coupled VHHanti-Spot-Tag single domain antibody (sdAb) fragment
ConjugateUnconjugated
Beads~ 90 µm (cross-linked 4 % agarose beads)
Binding capacity10 mg protein (30 kDa) / 1 mL settled resin (340 nmol / 1 mL settled resin)
Elution buffer0.1 mM Spot-peptide or 100 mM glycine pH 2.0 at 4°C
Storage ConditionShipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Shipped at ambient temperature. Upon receipt store at -20°C. Aliquot upon reconstitution and store at -20°C. Avoid repeated freeze / thaw cycles