ChromoTek Spot-peptide

Validation Data Gallery

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  Model fig of Spot peptide.

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  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.

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  Workflow of Spot-Cap purification.

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  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).

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  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.

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  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.

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  Application of Spot-tag related products.

Spot-peptide

Cat No. ep

Publications(1)

Peptide

Spot-Tag^®

Applications

Elution

Conjugate

Unconjugated

Synonyms

Spot peptide

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Product Information

Spot-peptide

• Applications: Elution of Spot-tagged fusion proteins bound to Spot-Cap
• Sequence: Peptide of amino acid sequence PDRVRAVSHWSS
• Form: Lyophilized powder
• Conjugate: Unconjugated
• Molecular weight: 1,397 g/mol
• Purity: >96% (HPLC)
• Reconstitution: Dissolve 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.
• Dilution: Dilute Spot-peptide to 0.1 mM in PBS for elution of Spot-tagged ; proteins from Spot-Cap.
• Storage Condition: Upon receipt store at -20°C/-4°F. ; Aliquot upon reconstitution and store at -20°C/-4°F. ; Avoid repeated freeze / thaw cycles.
• Shipment: Shipped at ambient temperature. Upon receipt store at -20°C. ; Aliquot upon reconstitution and store at -20°C. Avoid repeated ; freeze / thaw cycles.
• Spot-Tag® Origin: Engineered 12 amino acid sequence modified from the unstructured N-terminus of beta-catenin
• Cross Reactivity: The Spot-Nanobody shows minimal cross reactivity with beta-catenin.

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Publications

Application	Title
	Cells; Cep192, a Novel Missing Link between the Centrosomal Core and Corona in Dictyostelium Amoebae.; Authors - Valentin Pitzen