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ChromoTek Ubiquitin-Trap Agarose

The ChromoTek Ubiquitin-Trap Agarose consists of an anti-Ubiquitin Nanobody/VHH, which is coupled to agarose beads. It can be used for the immunoprecipitation of ubiquitin and ubiquitinylated proteins from mammalian, yeast, and plant cell extracts.

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 : uta

Synonyms

polyubiquitin B, UBB, ubiquitin, ubiquitin B

Validation Data Gallery

  • The ubiquitin-trap agarose (uta) 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 (uta) 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 (uta) 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 (uta) 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 ChromoTek Ubiquitin-Trap Agarose consists of an anti-Ubiquitin Nanobody/VHH, which is coupled to agarose beads. It can be used for the immunoprecipitation of ubiquitin and ubiquitinylated proteins from mammalian, yeast, and plant cell extracts.

DescriptionImmunoprecipitation of ubiquitin and ubiquitinylated proteins with anti-Ubiquitin Nanobody conjugated to agarose beads

• 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 (KD) 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!

Publications

ApplicationTitle
WB

mBio

A human papillomavirus 16 E2-TopBP1 dependent SIRT1-p300 acetylation switch regulates mitotic viral and human protein levels and activates the DNA damage response

Authors - Apurva T Prabhakar
View Article