使用敲除细胞株进行验证

Anti-HAUSP / USP7抗体(ab101648)

概述

  • 产品名称Anti-HAUSP / USP7抗体
    参阅全部 HAUSP / USP7 一抗
  • 描述
    兔多克隆抗体to HAUSP / USP7
  • 经测试应用适用于: WB, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Horse, Pig, Macaque Monkey
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human HAUSP/ USP7.

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; A431; MCF7; Ramos; PC3; as well as HeLa nuclear extract, Mouse Testis tissue lysate and Rat Testis tissue lysate.

性能

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应用

Our Abpromise guarantee covers the use of ab101648 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 140 kDa (predicted molecular weight: 128 kDa).
ICC/IF Use a concentration of 5 µg/ml.

靶标

  • 功能Hydrolase that deubiquitinates target proteins such as FOXO4, p53/TP53, MDM2, ERCC6, DNMT1, UHRF1, PTEN and DAXX (PubMed:11923872, PubMed:15053880, PubMed:16964248, PubMed:18716620, PubMed:25283148). Together with DAXX, prevents MDM2 self-ubiquitination and enhances the E3 ligase activity of MDM2 towards p53/TP53, thereby promoting p53/TP53 ubiquitination and proteasomal degradation. Deubiquitinates p53/TP53, preventing degradation of p53/TP53, and enhances p53/TP53-dependent transcription regulation, cell growth repression and apoptosis (PubMed:25283148). Deubiquitinates p53/TP53 and MDM2 and strongly stabilizes p53/TP53 even in the presence of excess MDM2, and also induces p53/TP53-dependent cell growth repression and apoptosis. Deubiquitination of FOXO4 in presence of hydrogen peroxide is not dependent on p53/TP53 and inhibits FOXO4-induced transcriptional activity. In association with DAXX, is involved in the deubiquitination and translocation of PTEN from the nucleus to the cytoplasm, both processes that are counteracted by PML. Involved in cell proliferation during early embryonic development. Involved in transcription-coupled nucleotide excision repair (TC-NER) in response to UV damage: recruited to DNA damage sites following interaction with KIAA1530/UVSSA and promotes deubiquitination of ERCC6, preventing UV-induced degradation of ERCC6. Contributes to the overall stabilization and trans-activation capability of the herpesvirus 1 trans-acting transcriptional protein ICP0/VMW110 during HSV-1 infection. Involved in maintenance of DNA methylation via its interaction with UHRF1 and DNMT1: acts by mediating deubiquitination of UHRF1 and DNMT1, preventing their degradation and promoting DNA methylation by DNMT1 (PubMed:21745816). Exhibits a preference towards 'Lys-48'-linked ubiquitin chains. Increases regulatory T-cells (Treg) suppressive capacity by deubiquitinating and stabilizing the transcription factor FOXP3 which is crucial for Treg cell function (PubMed:23973222).
  • 组织特异性Widely expressed. Overexpressed in prostate cancer.
  • 序列相似性Belongs to the peptidase C19 family.
    Contains 1 MATH domain.
    Contains 1 USP domain.
  • 结构域The C-terminus plays a role in its oligomerization.
  • 翻译后修饰Isoform 1: Phosphorylated. Isoform 1 is phosphorylated at positions Ser-18 and Ser-963. Isoform 2: Not phosphorylated.
    Isoform 1: Polyneddylated. Isoform 2: Not Polyneddylated.
    Isoform 1 and isoform 2: Not sumoylated.
    Isoform 1 and isoform 2: Polyubiquitinated by herpesvirus 1 trans-acting transcriptional protein ICP0/VMW110; leading to its subsequent proteasomal degradation. Isoform 1: Ubiquitinated at Lys-869.
  • 细胞定位Nucleus. Cytoplasm. Nucleus, PML body. Present in a minority of ND10 nuclear bodies. Association with ICP0/VMW110 at early times of infection leads to an increased proportion of USP7-containing ND10. Colocalizes with ATXN1 in the nucleus. Colocalized with DAXX in speckled structures. Colocalized with PML and PTEN in promyelocytic leukemia protein (PML) nuclear bodies.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Deubiquitinating enzyme 7 antibody
    • HAUSP antibody
    • Herpes virus associated ubiquitin specific protease antibody
    • Herpesvirus-associated ubiquitin-specific protease antibody
    • TEF 1 antibody
    • tef-1 antibody
    • TEF1 antibody
    • Ubiquitin carboxyl terminal hydrolase 7 antibody
    • Ubiquitin carboxyl-terminal hydrolase 7 antibody
    • Ubiquitin specific peptidase 7 (herpes virus associated) antibody
    • Ubiquitin specific peptidase 7 antibody
    • Ubiquitin specific peptidase 7 herpes virus associated antibody
    • Ubiquitin specific processing protease 7 antibody
    • Ubiquitin specific protease 7 (herpes virus associated) antibody
    • Ubiquitin specific protease 7 antibody
    • Ubiquitin specific protease 7 herpes virus associated antibody
    • Ubiquitin thioesterase 7 antibody
    • Ubiquitin thiolesterase 7 antibody
    • Ubiquitin-specific-processing protease 7 antibody
    • UBP 7 antibody
    • UBP-7 antibody
    • UBP7 antibody
    • UBP7_HUMAN antibody
    • USP 7 antibody
    • usp-7 antibody
    • Usp7 antibody
    • VMW110-ASSOCIATED PROTEIN, 135-KD antibody
    see all

Anti-HAUSP / USP7 antibody 图像



  • Predicted band size : 128 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: HAUSP/USP7 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: MCF7 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab101648 observed at 115 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab101648 was shown to recognize HAUSP/USP7 when HAUSP/USP7 knockout samples were used, along with additional cross-reactive bands. Wild-type and HAUSP/USP7 knockout samples were subjected to SDS-PAGE. ab101648 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/2000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-HAUSP / USP7 antibody (ab101648) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 7 : PC3 whole cell lysate (ab3954)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 128 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 68 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
  • ICC/IF image of ab101648 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab101648 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-HAUSP / USP7 antibody (ab101648) at 1 µg/ml

    Lane 1 : Testis (Mouse) Tissue Lysate
    Lane 2 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 128 kDa
    Observed band size : 128 kDa
    Additional bands at : 45 kDa,60 kDa,90 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

Anti-HAUSP / USP7 antibody (ab101648)参考文献

ab101648 has not yet been referenced specifically in any publications.

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