• Product name
  • Description
    Rabbit polyclonal to OTUB1
  • Host species
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Dog, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of Human OTUB1.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • Recombinant Human OTUB1 protein (ab87682) can be used as a positive control in WB. This antibody gave a positive signal in both Human and Mouse brain tissue lysate.



Our Abpromise guarantee covers the use of ab101471 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 31 kDa).
ICC/IF Use a concentration of 5 µg/ml.


  • Function
    Hydrolase that can remove conjugated ubiquitin from proteins and plays an important regulatory role at the level of protein turnover by preventing degradation. Regulator of T-cell anergy, a phenomenon that occurs when T-cells are rendered unresponsive to antigen rechallenge and no longer respond to their cognate antigen. Acts via its interaction with RNF128/GRAIL, a crucial inductor of CD4 T-cell anergy. Isoform 1 destabilizes RNF128, leading to prevent anergy. In contrast, isoform 2 stabilizes RNF128 and promotes anergy. Surprisingly, it regulates RNF128-mediated ubiquitination, but does not deubiquitinate polyubiquitinated RNF128. Deubiquitinates estrogen receptor alpha (ESR1). Mediates deubiquitination of 'Lys-48'-linked polyubiquitin chains, but not 'Lys-63'-linked polyubiquitin chains. Not able to cleave di-ubiquitin. Also capable of removing NEDD8 from NEDD8 conjugates, but with a nuch lower preference compared to 'Lys-48'-linked ubiquitin.
  • Tissue specificity
    Isoform 1 is ubiquitous. Isoform 2 is expressed only in lymphoid tissues such as tonsils, lymph nodes and spleen, as well as peripheral blood mononuclear cells.
  • Sequence similarities
    Belongs to the peptidase C65 family.
    Contains 1 OTU domain.
  • Domain
    In addition to ubiquitin-binding at the Cys-91 active site, a proximal ubiquitin-binding site is also present at Cys-23 Occupancy of the active site is needed to enable tight binding to the second site. Distinct binding sites for the ubiquitins may allow to discriminate among different isopeptide linkages (i.e. 'Lys-48'-, 'Lys-63'-linked polyubiquitin) in polyubiquitin substrates and achieve linkage-specific deubiquitination.
  • Cellular localization
  • Information by UniProt
  • Database links
  • Alternative names
    • Deubiquitinating enzyme OTUB1 antibody
    • hOTU1 antibody
    • HSPC263 antibody
    • OTB1 antibody
    • OTU domain containing ubiquitin aldehyde binding protein 1 antibody
    • OTU domain, ubiquitin aldehyde binding 1 antibody
    • OTU domain-containing Ubal-binding protein 1 antibody
    • OTU domain-containing ubiquitin aldehyde-binding protein 1 antibody
    • OTU-domain Ubal-binding 1 antibody
    • OTU1 antibody
    • Otub1 antibody
    • OTUB1_HUMAN antibody
    • Otubain 1 antibody
    • Otubain-1 antibody
    • Ubiquitin specific processing protease OTUB1 antibody
    • Ubiquitin thioesterase OTUB1 antibody
    • ubiquitin-specific protease otubain 1 antibody
    • Ubiquitin-specific-processing protease OTUB1 antibody
    see all


  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: OTUB1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Hek293 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab101471 observed at 31 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab101471 was shown to recognize OTUB1 in wild-type HAP1 cells as signal was lost at the expected MW in OTUB1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and OTUB1 knockout samples were subjected to SDS-PAGE. ab101471 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1µg/mL and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-OTUB1 antibody (ab101471) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 31 kDa
    Observed band size: 31 kDa
    Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.

    Exposure time: 12 minutes

    Secondary antibody - goat anti-rabbit HRP (ab97080)

  • ICC/IF image of ab101471 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab101471, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, goat anti-rabbit DyLight® 488 (IgG H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min)HeLa, HepG2 and MCF7 cells at 5µg/ml.


ab101471 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Flow Cytometry
Mouse Cell (spleen CD4 T cells)
Yes - Foxp3 Fixation/Permeabilization
Gating Strategy
viable cells
spleen CD4 T cells
Foxp3 Fixation/Permeabilization

Dr. Cinthia Stempin

Verified customer

Submitted Aug 03 2018

Abcam has not validated the combination of species/application used in this Abreview.
Human Cell lysate - whole cell (HEK293 cells)
Total protein in input
20 µg
Immuno-precipitation step
Protein A
HEK293 cells

Abcam user community

Verified customer

Submitted Dec 02 2013

Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Human Cell lysate - whole cell (HEK293 cells)
HEK293 cells
untreated or HA-OTUB1 transfected for 48 hours
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 12 2013


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