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使用敲除细胞株进行验证重组RabMAb

重组Anti-OTUB1抗体[EPR13028(B)] (ab175200)

Reviews (3) Submit a question References (13)
Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
  • Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
  • Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
  • Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
  • Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
  • Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-OTUB1抗体[EPR13028(B)]
    参阅全部 OTUB1 一抗

  • 描述

    兔单克隆抗体[EPR13028(B)] to OTUB1

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IPmore details
    不适用于: Flow Cyt,ICC/IF or IHC-P

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human OTUB1 aa 200 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
    Database link: Q96FW1

  • 阳性对照

    • WB: Wild-type HAP1, HeLa, MCF7, HepG2, HEK-293T, and HEK-293 cell lysates. Rat and mouse heart tissue lysates. IP: HeLa cell lysate.

  • 常规说明

    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
    For more information see here.

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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab175200于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (2)
1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
IP (1)
1/10 - 1/100.
说明
WB
1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
IP
1/10 - 1/100.
应用说明
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

靶标

  • 功能

    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.

  • 组织特异性

    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.

  • 序列相似性

    Belongs to the peptidase C65 family.
    Contains 1 OTU domain.

  • 结构域

    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.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
    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 kDa



    Lanes 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.

     

  • Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
    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 kDa



    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.

  • Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
    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)
    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 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunoprecipitation - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
    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.

  • Western blot - Anti-OTUB1 antibody [EPR13028(B)] (ab175200)
    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)
    Anti-OTUB1 antibody [EPR13028(B)] (ab175200)

文献 (13)

发表研究结果有使用 ab175200？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab175200 被引用在 13 文献中.

  • Shen J  et al. Deubiquitylating Enzyme OTUB1 Facilitates Neuronal Survival After Intracerebral Hemorrhage Via Inhibiting NF-κB-triggered Apoptotic Cascades. Mol Neurobiol 61:1726-1736 (2024). PubMed: 37775718
  • Zhu Q  et al. OTUB1 promotes osteoblastic bone formation through stabilizing FGFR2. Signal Transduct Target Ther 8:142 (2023). PubMed: 37024477
  • 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
  • Lin F  et al. Mesenchymal stem cells protect against ferroptosis via exosome-mediated stabilization of SLC7A11 in acute liver injury. Cell Death Dis 13:271 (2022). PubMed: 35347117
  • Henning NJ  et al. Deubiquitinase-targeting chimeras for targeted protein stabilization. Nat Chem Biol 18:412-421 (2022). PubMed: 35210618
View all Publications for this product

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-OTUB1 antibody [EPR13028(B)]

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HAP1 cell line)
Gel Running Conditions
Non-reduced Denaturing (gradient 4-12%)
Loading amount
50 µg
Specification
HAP1 cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 21 2017

Immunoprecipitation abreview for Anti-OTUB1 [EPR13028(B)] antibody

 Excellent
Abreviews
abreview image
Application
Immunoprecipitation
Immuno-precipitation step
Protein A
Sample
Human Cell lysate - whole cell (HEK293 cells)
Specification
HEK293 cells
Total protein in input
20 µg
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 12 2013

Western blot abreview for Anti-OTUB1 [EPR13028(B)] antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (HEK293 cells)
Specification
HEK293 cells
Treatment
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 12 2013

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