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Rabbit Recombinant Monoclonal OTUB1 antibody. Suitable for IP, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 3 publications.

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Images

Western blot - Anti-OTUB1 antibody [EPR24917-75] (AB270959), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-OTUB1 antibody [EPR24917-75] (AB270959), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-OTUB1 antibody [EPR24917-75] (AB270959), expandable thumbnail
  • Immunoprecipitation - Anti-OTUB1 antibody [EPR24917-75] (AB270959), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-OTUB1 antibody [EPR24917-75] (AB270959), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPFlow Cyt (Intra)WBIHC-PICC/IF
Human
Tested
Tested
Tested
Not recommended
Tested
Mouse
Expected
Expected
Tested
Not recommended
Expected
Rat
Expected
Expected
Tested
Not recommended
Expected

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/250
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Target data

Function

Hydrolase that can specifically remove 'Lys-48'-linked conjugated ubiquitin from proteins and plays an important regulatory role at the level of protein turnover by preventing degradation (PubMed:12401499, PubMed:12704427, PubMed:14661020, PubMed:23827681). 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 (PubMed:14661020). Acts via its interaction with RNF128/GRAIL, a crucial inductor of CD4 T-cell anergy (PubMed:14661020). Isoform 1 destabilizes RNF128, leading to prevent anergy (PubMed:14661020). In contrast, isoform 2 stabilizes RNF128 and promotes anergy (PubMed:14661020). Surprisingly, it regulates RNF128-mediated ubiquitination, but does not deubiquitinate polyubiquitinated RNF128 (PubMed:14661020). Deubiquitinates estrogen receptor alpha (ESR1) (PubMed:19383985). Mediates deubiquitination of 'Lys-48'-linked polyubiquitin chains, but not 'Lys-63'-linked polyubiquitin chains (PubMed:18954305, PubMed:19211026, PubMed:23827681). Not able to cleave di-ubiquitin (PubMed:18954305, PubMed:23827681). Also capable of removing NEDD8 from NEDD8 conjugates, but with a much lower preference compared to 'Lys-48'-linked ubiquitin (PubMed:18954305, PubMed:23827681). Plays a key non-catalytic role in DNA repair regulation by inhibiting activity of RNF168, an E3 ubiquitin-protein ligase that promotes accumulation of 'Lys-63'-linked histone H2A and H2AX at DNA damage sites (PubMed:20725033, PubMed:22325355). Inhibits RNF168 independently of ubiquitin thioesterase activity by binding and inhibiting UBE2N/UBC13, the E2 partner of RNF168, thereby limiting spreading of 'Lys-63'-linked histone H2A and H2AX marks (PubMed:20725033, PubMed:22325355). Inhibition occurs by binding to free ubiquitin: free ubiquitin acts as an allosteric regulator that increases affinity for UBE2N/UBC13 and disrupts interaction with UBE2V1 (PubMed:20725033, PubMed:22325355). The OTUB1-UBE2N/UBC13-free ubiquitin complex adopts a configuration that mimics a cleaved 'Lys48'-linked di-ubiquitin chain (PubMed:20725033, PubMed:22325355). Acts as a regulator of mTORC1 and mTORC2 complexes (PubMed:29382726, PubMed:35927303). When phosphorylated at Tyr-26, acts as an activator of the mTORC1 complex by mediating deubiquitination of RPTOR via a non-catalytic process: acts by binding and inhibiting the activity of the ubiquitin-conjugating enzyme E2 (UBE2D1/UBCH5A, UBE2W/UBC16 and UBE2N/UBC13), thereby preventing ubiquitination of RPTOR (PubMed:35927303). Can also act as an inhibitor of the mTORC1 and mTORC2 complexes in response to amino acids by mediating non-catalytic deubiquitination of DEPTOR (PubMed:29382726).

Alternative names

OTB1, OTU1, HSPC263, OTUB1, Ubiquitin thioesterase OTUB1, Deubiquitinating enzyme OTUB1, OTU domain-containing ubiquitin aldehyde-binding protein 1, Otubain-1, Ubiquitin-specific-processing protease OTUB1, hOTU1

Recommended products

Rabbit Recombinant Monoclonal OTUB1 antibody. Suitable for IP, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 3 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR24917-75
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Please note: IP, FC and ICC are valid for Human samples only

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The OTUB1 protein also known as OTU domain-containing ubiquitin aldehyde-binding protein 1 plays an important role in the removal of ubiquitin from proteins. This process is important for regulating protein stability and function within the cell. OTUB1 has a mass of approximately 32 kDa and is expressed ubiquitously across various tissues indicating its essential nature in cellular functions.

Biological function summary

OTUB1 functions as a deubiquitinating enzyme that cleaves ubiquitin from substrates influencing various cellular processes. It acts independently or as part of larger protein complexes. By controlling protein degradation pathways OTUB1 impacts protein turnover and cellular homeostasis. Interactions with other proteins allow OTUB1 to exert influence on different signaling pathways and cellular stress responses.

Pathways

OTUB1 holds a significant position in the regulation of the ubiquitin-proteasome system and DNA damage repair pathway. It closely interacts with proteins such as UBE2N affecting the DNA damage response. Additionally OTUB1 regulates processes through modulating pathways like NF-kB by interacting with and stabilizing specific protein factors involved.

Associated diseases and disorders

OTUB1’s function connects it to various pathological states like cancer and neurodegenerative diseases. Aberrant expression or mutations in OTUB1 can lead to disruptions in cell cycle regulation and apoptosis contributing to oncogenesis. In the context of neurodegenerative diseases OTUB1 is related to tau stabilization with proteins such as HSP90 playing a role in the same regulatory mechanisms highlighting its relevance in these disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Lanes 1-3: Merged signal (red and green). Green -ab270959 observed at 31kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa. ab270959 Anti-OTUB1 antibody [EPR24917-75] was shown to specifically react with OTUB1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human OTUB1 knockout HEK-293T cell line ab266551 (knockout cell lysate Human OTUB1 knockout HEK-293T cell lysate ab257569) was used. Wild-type and OTUB1 knockout samples were subjected to SDS-PAGE.

    ab270959 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959) at 1/1000 dilution

    Lane 1: Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 2: OTUB1 knockout HEK293T (Human OTUB1 knockout HEK-293T cell line ab266551) whole cell lysate at 20 µg

    Lane 2: Western blot - Human OTUB1 knockout HEK-293T cell line (Human OTUB1 knockout HEK-293T cell line ab266551)

    Lane 3: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution

    Predicted band size: 31 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling OTUB1 with ab270959 at 1/250 (2.204 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in HeLa cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized OTUB1 KO HEK293T (Human OTUB1 knockout HEK-293T cell line ab266551) cells labelling OTUB1 with ab270959 at 1/250 (2.204 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing no staining in OTUB1 KO HEK293T cell line and nuclear and cytoplasmic staining in Parental HEK293T is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

  • Immunoprecipitation - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Immunoprecipitation - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    OTUB1 was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate 10ug with ab270959 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270959 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate 10ug

    Lane 2: ab270959 IP in HeLa whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab270959 in HeLa whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 7 seconds

    All lanes: Immunoprecipitation - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Predicted band size: 31 kDa

  • Flow Cytometry (Intracellular) - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HEK293T (human embryonic kidney epithelial cell, Right)/ OTUB1 KO HEK293T (Left) cells labelling OTUB1 with ab270959 at 1/500 dilution (0.1ug)/ (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Positive staining on 293T cells (Human wild-type HEK-293T cell line ab255449), while no staining on OTUB1 knockout HEK-293T cells (Human OTUB1 knockout HEK-293T cell line ab266551).

  • Flow Cytometry (Intracellular) - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling OTUB1 with ab270959 at 1/500 dilution (0.1ug)/ (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 26 seconds

    All lanes: Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959) at 1/1000 dilution

    Lane 1: MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 2: U-2 OS(human bone osteosarcoma epithelial cell), whole cell lysate at 20 µg

    Lane 3: Mouse liver tissue lysate at 20 µg

    Lane 4: Mouse brain tissue lysate at 20 µg

    Lane 5: Rat liver tissue lysate at 20 µg

    Lane 6: Rat brain tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 31 kDa

  • Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959), expandable thumbnail

    Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 15 seconds

    All lanes: Western blot - Anti-OTUB1 antibody [EPR24917-75] (ab270959) at 1/1000 dilution

    Lane 1: C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

    Lane 2: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

    Lane 3: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

    Lane 4: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 31 kDa

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Product protocols

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