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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

Validation Data Gallery

  • 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 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.

    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.

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.

DescriptionThe 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.

ApplicationsIP, CoIP
Specificity/TargetBinds 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 ReactivityHuman, Mouse, Hamster, Dog, Spinach, Yeast
ConjugateAgarose beads; ~90 um (cross-linked 4% agarose beads)
Elution buffer2x SDS-sample buffer (Lammli)
Wash Buffer Compatibility2M NaCl, 5 mM DTT, 0 mM β-mercaptoethanol, 5 mM TCEP, 2% NP40, 2% Triton X-100, 0% SDS, 2-3 M Urea
TypeNanobody
ClassRecombinant
HostCamelid
Affinity The KD for monomeric ubiquitin is 90 nM. The apparent affinity is even higher for ubiquitin chains due to avidity effects.
Compatibility with mass spectrometryThe Ubiquitin-Trap is optimized for on-bead digestion. For the application note, please click here: On-bead digest protocol for mass spectrometry
RRIDAB_2943577
Storage Buffer20% ethanol
Storage ConditionShipped at ambient temperature. Upon receipt store at +4°C. Stable for one year. DO not freeze!

Kit components

Component Description
Ubiquitin-Trap Agarose20 reactions (500 µl)
Lysis bufferOptimized for cytoplasmic proteins and mammalian cell lysis
RIPA bufferOptimized for nuclear/chromatin proteins and mammalian cell lysis
Wash bufferRemoval of unwanted proteins, peptides, etc.
Dilution bufferDilution of cell lysate
Elution bufferFor acidic elution