概述

  • 产品名称Anti-USP9x抗体
    参阅全部 USP9x 一抗
  • 描述
    兔多克隆抗体to USP9x
  • 特异性ab19879 detects 289kDa full length USP9X Human protein (Q93008) in WB on Caco2 Lysate. All detected bands are quenched by the immunizing peptide ab20617
  • 经测试应用适用于: IP, WB, IHC (PFA fixed), ICC/IF, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human USP9x.

  • 阳性对照
    • USP9x can be detected in the polarized intestinal cell line T84 which is a human colon carcinoma epithelial cell line. In tissue culture: USP9x is also observed in L-cell fibroblasts and MDCK cells. USP9x is ubiquitously expressed in all adult tissues.

性能

相关产品

应用

Our Abpromise guarantee covers the use of ab19879 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
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100,105, 290 kDa (predicted molecular weight: 54, 100,105 , 290 kDa).
IHC (PFA fixed) Use a concentration of 0.1 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.

靶标

  • 功能Deubiquitinase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. May therefore play an important role regulatory role at the level of protein turnover by preventing degradation of proteins through the removal of conjugated ubiquitin. Essential component of TGF-beta/BMP signaling cascade. Regulates chromosome alignment and segregation in mitosis by regulating the localization of BIRC5/survivin to mitotic centromeres. Specifically hydrolyzes both 'Lys-29'- and 'Lys-33'-linked polyubiquitins chains. Specifically deubiquitinates monoubiquitinated SMAD4, opposing the activity of E3 ubiquitin-protein ligase TRIM33.
  • 组织特异性Widely expressed in embryonic and adult tissues.
  • 序列相似性Belongs to the peptidase C19 family.
  • 细胞定位Cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Deubiquitinating enzyme FAF X antibody
    • Deubiquitinating enzyme FAF-X antibody
    • DFFRX antibody
    • Drosophila fat facets related X linked antibody
    • FAF antibody
    • Fafl antibody
    • Fam antibody
    • Fat facets homolog antibody
    • Fat facets in mammals antibody
    • Fat facets protein related X linked antibody
    • Fat facets protein related, X-linked antibody
    • Fat facets protein-related antibody
    • hFAM antibody
    • MRX99 antibody
    • Probable ubiquitin carboxyl terminal hydrolase FAF X antibody
    • Probable ubiquitin carboxyl-terminal hydrolase FAF-X antibody
    • Ubiquitin carboxyl-terminal hydrolase FAM antibody
    • Ubiquitin specific peptidase 9 X linked antibody
    • Ubiquitin specific peptidase 9, X-linked antibody
    • Ubiquitin specific processing protease FAF X antibody
    • Ubiquitin specific protease 9 X chromosome antibody
    • Ubiquitin thioesterase FAF X antibody
    • Ubiquitin thiolesterase FAF X antibody
    • Ubiquitin thiolesterase FAF-X antibody
    • Ubiquitin-specific protease 9 antibody
    • Ubiquitin-specific-processing protease FAF-X antibody
    • USP9 (gene name) antibody
    • Usp9x antibody
    • USP9X_HUMAN antibody
    • Uubiquitin specific protease 9, X chromosome (fat facets like Drosophila) antibody
    • X chromosome antibody
    • X-linked antibody
    see all

Anti-USP9x antibody 图像



  • Predicted band size : 54, 100,105 , 290 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: USP9x knockout HAP1 cell lysate (20 µg)
    Lane 3: T84 cell lysate (20 µg)
    Lane 4: NIH3T3 cell lysate (20 µg)
    Lanes 1 to 4: Merged signal (red and green). Green - ab19879 observed at 290 kDa. Red - loading control, ab181602 observed at 124 kDa.
    ab19879 was shown to specifically react with USP9x when USP9x knockout samples were used. Wild-type and USP9x knockout samples were subjected to SDS-PAGE. ab9879 and ab181602 (loading control to GAPDH) were both diluted at 1 µg/ml and 1/10 000 respectively 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.


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 54, 100,105 , 290 kDa
    Observed band size : 100,105,290 kDa (why is the actual band size different from the predicted?)
    Additional bands at : ~50-65 kDa. We are unsure as to the identity of these extra bands.

    ab19879 detects full length USP9x protein as well as a number of  USP9x fragments in WB on Caco2 Lysate:

    289kDa Human protein: Q93008 USP9X  (Full length protein)
    105kDa Human protein: Q6P468 - USP9X protein (Fragment) Human protein 
    99.7kDa Human protein: Q59EZ5 - USP9X protein variant (Fragment).
    53.9kDa Q86X58 - USP9X protein (Fragment).

  • Immuofluorescent staining for USP9X in the rat hippocampus (dentate gyrus) using ab19879 (1/300 = 0.07µg/ml). Image is taken with X10 objective. ab19879 was incubated overnight at RT. Secondary antibody used was anti-rabbit Alexa fluor 488 (1/1000 for 2h at RT). Rats were intracardially perfused with paraformaldehyde 4%, brain tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut on a cryostat for free floating IHC.

  • ICC/IF image of ab19879 stained Hek293 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 (ab19879, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
  • USP9x was immunoprecipitated using 0.5mg Caco2 whole cell extract, 5µg of Rabbit polyclonal to USP9x and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Caco2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19879.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 290kDa: USP9x.

Anti-USP9x antibody (ab19879)参考文献

This product has been referenced in:
  • Agrawal P  et al. Ubiquitin Specific Peptidase 9, X-linked (USP9X) Modulates the Activity of Mammalian Target of Rapamycin (mTOR). J Biol Chem : (2012). IP ; Human . Read more (PubMed: 22544753) »
  • Marx C  et al. ErbB2 trafficking and degradation associated with K48 and K63 polyubiquitination. Cancer Res 70:3709-17 (2010). WB ; Human . Read more (PubMed: 20406983) »

See all 3 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Sections of rat brain)
Specification Sections of rat brain
Fixative Paraformaldehyde
Username

Dr. Sophie Pezet

Verified customer

提交于 May 18 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"