ChromoTek iST Spot-Trap Kit for AP-MS sample preparation of Spot-fusion proteins

Kit for IP/ Co-IP of Spot-tagged proteins & sample preparation for mass spectrometry (MS)







Cat no : etak-iST


Spot tag, Spot, PDRVRAVSHWSS, Spot-Trap

Product Information

Kit for IP/ Co-IP of Spot-tagged proteins & sample preparation for mass spectrometry (MS)

DescriptionThe iST Spot-Trap® Kit enables researchers to process Spot-tagged proteins and their interacting partners for Mass Spectrometric analysis. The kit includes the ChromoTek Spot-Trap for immunoprecipitation/Co-IP of Spot-tagged proteins and the PreOmics iST buffers and cartridges required for bottom-up proteomic sample preparation. This robust method yields purified peptides while dramatically reducing contamination and sample loss. Each kit accommodates up to eight samples and includes pull-down reagent for controls:

• Reliable dentification of Spot-tagged proteins & interacting partners by mass spectrometry.

• Identification of even low abundant/ low expressed Spot-tagged proteins

• Superior processing of IP samples into clean peptide mixtures

• Intuitive protocol and reproducible results

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)
Type Nanobody
Affinity (KD) Dissociation constant KD of 6 nM
Compatibility with mass spectrometryThe Spot-Trap® is optimized for on-bead digestion.
Storage Buffer20% ethanol
Storage ConditionKit is shipped at ambient temperature. Upon receipt store Spot-Trap at 4°C and Digest at -20°C
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

Spot-Trap Agarose8 reactions plus 2 controls
DigestEnzyme Trypsin-mix to digest proteins.
Resuspend bufferProtease reconstitution buffer for enzymes.
Lyse bufferDenature, reduce and alkylate proteins.
Stop solutionStop the enzymatic activity.
Wash 1 bufferClean up peptides from hydrophobic contaminants.
Wash 2 bufferClean up peptides from hydrophilic contaminants.
Elution bufferElute the peptides from the cartridge.
LC-Load loadLoad peptides on reversed-phase LC-MS column.
CartridgesCartridge for 1 to 100 µg protein starting material.
Waste tubesTube for collecting waste after washing steps.
Collection tubesTube for collecting peptides after elution.
AdaptersEnables placing a cartridge into a tube.
CapsCap to optionally close the cartridge’s bottom.