ChromoTek Ubiquitin-Trap Agarose Kit

The Ubiquitin-Trap Agarose Kit is a ready-to-use reagent for the IP of ubiquitin and ubiquitinylated proteins. It consists of an anti-ubiquitin Nanobody/VHH coupled to agarose beads, along with lysis, wash, and elution buffers to use in the IP process.

Specificity

monomeric ubiquitin, ubiquitin chains, and ubiquitinylated proteins

Reactivity

Pan-reactive. Tested with human, mouse, hamster, dog, spinach, yeast ubiquitin

Applications

IP, CoIP

Type

Nanobody

Conjugate

Agarose

Cat no : utak

Synonyms

polyubiquitin B, UBB, ubiquitin, ubiquitin B

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Validation Data Gallery

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  The ubiquitin-trap agarose kit (utak) was used to immunoprecipitate endogenous ubiquitin and ubiquitinylated proteins from human (HEK293T), mouse (C2C12), and hamster (CHO) cell lines treated with MG-132. For each IP, samples of the input lysate (I), non-bound flow-through (F), and bound (B) fractions were analyzed using western blot. Ubiquitin recombinant antibody (80992-1-RR) and HRP-conjugated Affinipure Goat Anti-Rabbit IgG (H+L) (SA00001-2) were used in the western blot analysis.

• 

  The ubiquitin-trap agarose kit (utak) was used to immunoprecipitate endogenous ubiquitin and ubiquitinylated proteins from dog (MDCK) cells, spinach (Spinacia oleracea), and baker's yeast (Saccharomyces cerevisiae) treated with MG-132. For each IP, samples of the input lysate (I), non-bound flow-through (F), and bound (B) fractions were analyzed using western blot. Ubiquitin recombinant antibody (80992-1-RR) and HRP-conjugated Affinipure Goat Anti-Rabbit IgG (H+L) (SA00001-2) were used in the western blot analysis.

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Product Information

The Ubiquitin-Trap Agarose Kit is a ready-to-use reagent for the IP of ubiquitin and ubiquitinylated proteins. It consists of an anti-ubiquitin Nanobody/VHH coupled to agarose beads, along with lysis, wash, and elution buffers to use in the IP process.

• Description: The Ubiquitin-Trap Agarose Kit contains Ubiquitin-Trap agarose, lysis, wash, and elution buffers for efficient immunoprecipitation of ubiquitin and ubiqutinylated proteins.; ;

  • Fast, reliable & efficient one-step immunoprecipitation;

  • Ready-to-use;

  • No heavy & light antibody chains;

  • Stable under harsh washing conditions;

  • Suitable for downstream mass spec analysis;

  • Works in samples from: mammals, plants, yeast, etc.

• Applications: IP, CoIP
• Specificity/Target: Binds to monomeric ubiquitin, ubiquitin chains, and ubiquitinylated proteins.; Ubiquitin chains may be linked via lysines; linkages via lysines such as K11, K48 and K63 are compatible with binding by the Ubiquitin-Trap (further linkage types also likely to be compatible, but not tested).
• Tested Reactivity: Human, Mouse, Hamster, Dog, Spinach, Yeast
• Conjugate: Agarose beads; ~90 um (cross-linked 4% agarose beads)
• Elution buffer: 2x SDS-sample buffer (Lammli)
• Wash Buffer Compatibility: 2M NaCl, 5 mM DTT, 0 mM β-mercaptoethanol, 5 mM TCEP, 2% NP40, 2% Triton X-100, 0% SDS, 2-3 M Urea
• Type: Nanobody
• Class: Recombinant
• Host: Camelid
• Affinity: The K_D for monomeric ubiquitin is 90 nM. The apparent affinity is even higher for ubiquitin chains due to avidity effects.
• Compatibility with mass spectrometry: The Ubiquitin-Trap is optimized for on-bead digestion. For the application note, please click here: On-bead digest protocol for mass spectrometry
• RRID: AB_2943577
• Storage Buffer: 20% ethanol
• Storage Condition: Shipped at ambient temperature. Upon receipt store at +4°C. Stable for one year. DO not freeze!

Kit components

Component	Description
Ubiquitin-Trap Agarose	20 reactions (500 µl)
Lysis buffer	Optimized for cytoplasmic proteins and mammalian cell lysis
RIPA buffer	Optimized for nuclear/chromatin proteins and mammalian cell lysis
Wash buffer	Removal of unwanted proteins, peptides, etc.
Dilution buffer	Dilution of cell lysate
Elution buffer	For acidic elution

Other Formats

Ubiquitin-Trap Agarose