Product nameAnti-OTUB1 antibody [EPR13028(B)]
See all OTUB1 primary antibodies
DescriptionRabbit monoclonal [EPR13028(B)] to OTUB1
Tested applicationsSuitable for: WB, IPmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human OTUB1 aa 200 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
Database link: Q96FW1
- WB: Wild-type HAP1, HeLa, MCF7, HepG2, HEK-293T, and HEK-293 cell lysates. Rat and mouse heart tissue lysates. IP: HeLa cell lysate.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab175200 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 31 kDa.|
|IP||1/10 - 1/100.|
FunctionHydrolase 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 specificityIsoform 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 similaritiesBelongs to the peptidase C65 family.
Contains 1 OTU domain.
DomainIn 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.
- Information by UniProt
- Deubiquitinating enzyme OTUB1 antibody
- hOTU1 antibody
- HSPC263 antibody
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: HEK-293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab175200 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab175200 was shown to specifically react with OTUB1 in wild type cells as signal was lost in OTUB1 knockout cells. Wild-type and OTUB1 knockout samples were subjected to SDS-PAGE. ab175200 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution 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.
ab175200 (purified) at 1:50 dilution (2ug) immunoprecipitating OTUB1 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab175200 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab175200 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes : Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Mouse heart whole tissue lysates
Lane 3 : Rat heart whole tissue lysates
Lysates/proteins at 15 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 31 kDa
Blocking and diluting buffer: 5% NFDM/TBST
Western blot analysis on immunoprecipitation pellet from MCF7 cell lysate labeling OTUB1 with unpurified ab175200 at 1/10 dilution.
All lanes : Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : 293T cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 31 kDa