Recombinant Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
![Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-388730-anti-otub1-antibody-epr13028b-western-blot-hek293t-lysates.jpg "Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13028(B)] to OTUB1
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-OTUB1 antibody [EPR13028(B)]
See all OTUB1 primary antibodies -
Description
Rabbit monoclonal [EPR13028(B)] to OTUB1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human OTUB1 aa 200 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
Database link: Q96FW1 -
Positive control
- WB: Wild-type HAP1, HeLa, MCF7, HepG2, HEK-293T, and HEK-293 cell lysates. Rat and mouse heart tissue lysates. IP: HeLa cell lysate.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR13028(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (2) |
1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
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| IP | (1) |
1/10 - 1/100.
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| Notes |
|---|
|
WB
1/1000 - 1/10000. Predicted molecular weight: 31 kDa. |
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IP
1/10 - 1/100. |
Target
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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
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 55611 Human
- Entrez Gene: 107260 Mouse
- Entrez Gene: 293705 Rat
- Omim: 608337 Human
- SwissProt: Q96FW1 Human
- SwissProt: Q7TQI3 Mouse
- SwissProt: B2RYG6 Rat
- Unigene: 473788 Human
see all -
Alternative names
- Deubiquitinating enzyme OTUB1 antibody
- hOTU1 antibody
- HSPC263 antibody
see all
Images
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Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)All lanes : Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : OTUB1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 31 kDaLanes 1-2: Merged signal (red and green). Green - ab175200 observed at 31 kDa. Red - loading control ab8245 observed at 37 kDa.
ab175200 Anti-OTUB1 antibody [EPR13028(B)] was shown to specifically react with OTUB1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266551 (knockout cell lysate ab257569) was used. Wild-type and OTUB1 knockout samples were subjected to SDS-PAGE. ab175200 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°CC at 1 in 1000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-285155-anti-otub1-antibody-epr13028b-western-blot.jpg)
Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)All lanes : Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : OTUB1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK-293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 31 kDaLanes 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.
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![Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-287376-anti-otub1-antibody-epr13028b-immunoprecipitation.jpg)
Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)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. -
![Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-286069-anti-otub1-antibody-epr13028b-western-blot.jpg)
Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)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.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 31 kDaBlocking and diluting buffer: 5% NFDM/TBST
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![Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-201692-anti-otub1-antibody-epr13028b-immunoprecipitation.jpg)
Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)Western blot analysis on immunoprecipitation pellet from MCF7 cell lysate labeling OTUB1 with unpurified ab175200 at 1/10 dilution.
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![Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-201691-anti-otub1-antibody-epr13028b-western-blot.jpg)
Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)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 -
![Anti-OTUB1 antibody [EPR13028(B)] (ab175200)](https://www.abcam.com/ps/products/175/ab175200/Images/ab175200-7-benefits-of-recombinant-antibodies.png)
Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab175200 has been referenced in 10 publications.
- Li D et al. CST1 inhibits ferroptosis and promotes gastric cancer metastasis by regulating GPX4 protein stability via OTUB1. Oncogene 42:83-98 (2023). PubMed: 36369321
- Henning NJ et al. Deubiquitinase-targeting chimeras for targeted protein stabilization. Nat Chem Biol 18:412-421 (2022). PubMed: 35210618
- Ling X et al. Ovarian tumorB1-mediated heat shock transcription factor 1 deubiquitination is critical for glycolysis and development of endometriosis. iScience 25:105363 (2022). PubMed: 36339263
- Xie P et al. The deubiquitinase OTUB1 fosters papillary thyroid carcinoma growth through EYA1 stabilization. J Cell Mol Med 25:10980-10989 (2021). PubMed: 34773364
- Yang L et al. Broad Spectrum Deubiquitinase Inhibition Induces Both Apoptosis and Ferroptosis in Cancer Cells. Front Oncol 10:949 (2020). PubMed: 32596160
- Xu Y et al. Anti-bacterial and anti-viral nanchangmycin displays anti-myeloma activity by targeting Otub1 and c-Maf. Cell Death Dis 11:818 (2020). PubMed: 32999280
- Liang C et al. LncRNA GAS6-AS2 facilitates tumor growth and metastasis of hepatocellular carcinoma by activating the PI3K/AKT/FoxO3a signaling pathway. Int J Clin Exp Pathol 12:4011-4023 (2019). PubMed: 31933797
- Zhou X et al. The deubiquitinase Otub1 controls the activation of CD8+ T cells and NK cells by regulating IL-15-mediated priming. Nat Immunol 20:879-889 (2019). PubMed: 31182807
- Liu T et al. The Deubiquitylase OTUB1 Mediates Ferroptosis via Stabilization of SLC7A11. Cancer Res 79:1913-1924 (2019). PubMed: 30709928
- Pasupala N et al. OTUB1 non-catalytically stabilizes the E2 ubiquitin-conjugating enzyme UBE2E1 by preventing its autoubiquitination. J Biol Chem 293:18285-18295 (2018). PubMed: 30282802