Immunoprecipitation of Spot-tagged protein.

Spot-Trap Magnetic Agarose for immunoprecipitation of Spot-tagged proteins: Coomassie blue stained gel and Western blot I: Input, FT: Flow-Through, B: Bound.

Application of Spot-tag related products.

Immunoprecipitation of Spot-tagged protein.

Immunoprecipitation of Spot-tagged protein. Input (I), non-bound (FT) and bound (B), Coomassie staining and Western blotting. Western blot indicates high effectivity.

Elution of Spot-tag fusion protein. Input (I), non-bound (FT) and eluted (E1, E2 and E3) fractions. Elutions with Spot-peptide.

Application of Spot-tag related products.

Make the most of your time & gain about 40 hours with the iST_Spot-Trap Kit.

Schematic workflow of iST_Spot-Tap_kit.

Preomics Technology Priciple.

Immunoprecipitation of Spot-tagged protein.

Immunoprecipitation of Spot-tagged protein. Input (I), non-bound (FT) and bound (B), Coomassie staining and Western blotting. Western blot indicates high effectivity.

Elution of Spot-tag fusion protein. Input (I), non-bound (FT) and eluted (E1, E2 and E3) fractions. Elutions with Spot-peptide.

Application of Spot-tag related products.

Immunoprecipitation of Spot-tagged protein.

Spot-Trap Magnetic Agarose for immunoprecipitation of Spot-tagged proteins: Coomassie blue stained gel and Western blot I: Input, FT: Flow-Through, B: Bound.

Application of Spot-tag related products.

Spot-Trap Magnetic Particles M-270 pull-down.

Immunoprecipitation (IP) of GFP with Spot-Trap Magnetic Particles M-270: Coomassie and Western blot (I: Input, FT: Flow-Through, B: Bound).

Application of Spot-tag related products.

Cartoon to visualize the binding of large protein with Spot-tag (Spot-tag (green) + protein of interest (POI, blue) with interacting partner X (Prot X, orange) or multimeric proteins (tag (green) + protein of interest (POI, blue) to Spot-Nanobody (dark green) of Spot-Trap Magnetic Agarose or Spot-Trap Magnetic Particles M-270.

Spot-Trap Magnetic Particles M-270 pull-down.

Immunoprecipitation (IP) of GFP with Spot-Trap Magnetic Particles M-270: Coomassie and Western blot (I: Input, FT: Flow-Through, B: Bound).

Application of Spot-tag related products.

Cartoon to visualize the binding of large protein with Spot-tag (Spot-tag (green) + protein of interest (POI, blue) with interacting partner X (Prot X, orange) or multimeric proteins (tag (green) + protein of interest (POI, blue) to Spot-Nanobody (dark green) of Spot-Trap Magnetic Agarose or Spot-Trap Magnetic Particles M-270.

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.

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.

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.