Rabbit Recombinant Monoclonal OTUB1 antibody. Suitable for IP, WB and reacts with Human, Mouse, Rat samples. Cited in 13 publications.
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IP | Flow Cyt | WB | IHC-P | ICC/IF | |
---|---|---|---|---|---|
Human | Tested | Not recommended | Tested | Not recommended | Not recommended |
Mouse | Expected | Not recommended | Tested | Not recommended | Not recommended |
Rat | Expected | Not recommended | Tested | Not recommended | Not recommended |
Species | Dilution info | Notes |
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Species Human | Dilution info 1/10 - 1/100 | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/10000 | Notes - |
Species Rat | Dilution info 1/1000 - 1/10000 | Notes - |
Species Human | Dilution info 1/1000 - 1/10000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
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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).
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
Rabbit Recombinant Monoclonal OTUB1 antibody. Suitable for IP, WB and reacts with Human, Mouse, Rat samples. Cited in 13 publications.
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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:
For more information, read more on recombinant antibodies.
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.
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.
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.
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.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Terms & Conditions.
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 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. ab175200 and Anti-GAPDH antibody [6C5] - Loading Control (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 (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 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: OTUB1 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human OTUB1 knockout HEK-293T cell line (Human OTUB1 knockout HEK-293T cell line ab266551)
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 130 kDa, 31 kDa
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab175200 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
Predicted band size: 31 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: OTUB1 knockout HAP1 whole cell lysate at 20 µg
Lane 3: HeLa whole cell lysate at 20 µg
Lane 4: HEK-293 whole cell lysate at 20 µg
Predicted band size: 31 kDa
Western blot analysis on immunoprecipitation pellet from MCF7 cell lysate labeling OTUB1 with unpurified ab175200 at 1/10 dilution.
All lanes: Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
Predicted band size: 31 kDa
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Lane 2: Mouse heart whole tissue lysates at 15 µg
Lane 3: Rat heart whole tissue lysates at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 31 kDa
All lanes: Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200) at 1/1000 dilution
Lane 1: HeLa cell lysate at 10 µg
Lane 2: HepG2 cell lysate at 10 µg
Lane 3: 293T cell lysate at 10 µg
Lane 4: MCF7 cell lysate at 10 µg
Predicted band size: 31 kDa
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