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Validation Data Gallery

  • Immunoprecipitation of Spot-tagged protein.

    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.

    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.

    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.

    Application of Spot-tag related products.

ChromoTek Spot-Trap® Agarose, Kit for Immunoprecipitation

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. etak

Specificity

Spot-Tag®

Applications

IP, CoIP, ChIP, RIP

Type

Nanobody

Conjugate

Agarose

Synonyms

Spot tag, Spot, PDRVRAVSHWSS, Spot-Trap

Formats:  Agarose kit
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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.

DescriptionThe Spot-Trap Agarose Kit contains Spot-Trap Agarose, lysis, wash, and elution buffers for efficient immunoprecipitation of Spot-tagged proteins and their interacting factors.

• No contaminating heavy and light antibody chains

• Ready-to-use

• Recombinantly expressed without batch-to-batch variation

• Well-characterized and validated

• Compatible with harsh buffer conditions

• Extraordinary low background compared to other affinity resins

• Short incubation time of 30-60 minutes

ApplicationsIP, CoIP, ChIP, RIP
Specificity/TargetSpot-Tag® sequence PDRVRAVSHWSS
Binding capacity17.5 μg of recombinant Spot-tagged protein (~30 kDa) per 25 μL bead slurry
ConjugateAgarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads)
Elution bufferSDS sample buffer
10 mM NaOH pH 12
Wash buffer compatibility2 M urea, 2 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100, 0.1 % SDS
Type Nanobody
ClassRecombinant
HostAlpaca
Affinity (KD) Dissociation constant KD of 6 nM
Compatibility with mass spectrometryThe Spot-Trap® is optimized for on-bead digestion. For the application note, please click here:
On-bead digest protocol for mass spectrometry
RRIDAB_2827590
Storage Buffer20% ethanol
Storage ConditionShipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Do not freeze!
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.

Kit components

ComponentDescription
Spot-Trap® Agarose20 reactions (500 µL)
Lysis bufferOptimized for cytoplasmatic proteins and mammalian cell lysis
RIPA bufferOptimized for nuclear/chromatin proteins and mammalian cell lysis
Wash buffer Removal of unwanted proteins, peptides, etc.
Dilution bufferDilution of cell lysate