ChromoTek Spot-Trap® Agarose

Validation Data Gallery

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  Immunoprecipitation of Spot-tagged protein.

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

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

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  Application of Spot-tag related products.

Spot-Trap® Agarose is an affinity bead for immunoprecipitation (IP) of Spot-tagged proteins It comprises an anti-Spot-Tag® VHH/ Nanobody coupled to agarose beads.

Cat No. eta

Publications(6)

Specificity

Spot-Tag^®

Applications

IP, CoIP, ChIP, RIP

Type

Nanobody

Conjugate

Agarose

Synonyms

Spot tag, Spot, PDRVRAVSHWSS, Spot-Trap

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Product Information

Spot-Trap® Agarose is an affinity bead for immunoprecipitation (IP) of Spot-tagged proteins It comprises an anti-Spot-Tag® VHH/ Nanobody coupled to agarose beads.

• Description: Immunoprecipitation of Spot-tagged proteins and their interacting factors with anti-Spot Nanobody conjugated to agarose beads.

  • No contaminating heavy and light antibody chains

  • Ready-to-use

  • Recombinantly expressed without batch-to-batch variation

  • Well-characterized and validated

  • High stability allows for stringent washing

  • Extraordinary low background compared to other affinity resins

  • Short incubation time of 30-60 minutes

• Applications: IP, CoIP, ChIP, RIP
• Specificity/Target: Spot-Tag sequence PDRVRAVSHWSS
• Binding capacity: 17.5 μg of recombinant Spot-tagged protein (~30 kDa) per 25 μL bead slurry
• Conjugate: Agarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads)
• Elution buffer: SDS sample buffer; 10 mM NaOH pH 12
• Wash buffer compatibility: 2 M urea, 2 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100, 0.1 % SDS
• Type: Nanobody
• Class: Recombinant
• Host: Alpaca
• Affinity (KD): Dissociation constant K_D of 6 nM
• Compatibility with mass spectrometry: The Spot-Trap® is optimized for on-bead digestion. For the application note, please click here:; On-bead digest protocol for mass spectrometry
• RRID: AB_2827590
• Storage Buffer: 20% ethanol
• Storage Condition: Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Do not freeze!
• 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.
• Size: 25ul/reactions (eg：20rxns=500ul slurry)

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Publications

Application	Title
IP	J Biol Chem; Serine phosphorylation of the small phosphoprotein ICAP1 inhibits its nuclear accumulation.; Authors - Valerie L Su
IP	Biomolecules; Nanobodies Right in the Middle: Intrabodies as Toolbox to Visualize and Modulate Antigens in the Living Cell.; Authors - Teresa R Wagner
	J Cell Sci; 30 years of nanobodies - an ongoing success story of small binders in biological research; Authors - Desiree I Frecot
	J Cell Sci; Identification of distinct active pools of yeast serine palmitoyltransferase in sub-compartments of the ER; Authors - Bianca M Esch
	Nature; TDP-43 forms amyloid filaments with a distinct fold in type A FTLD-TDP; Authors - Diana Arseni
IP	EMBO J; Jacob-induced transcriptional inactivation of CREB promotes Aβ-induced synapse loss in Alzheimer's disease; Authors - Katarzyna M Grochowska