Product Details

Uniprot No.

Target Names

Alternative Names

Probable ubiquitin carboxyl-terminal hydrolase FAF-X (EC 3.4.19.12) (Deubiquitinating enzyme FAF-X) (Fat facets in mammals) (hFAM) (Fat facets protein-related, X-linked) (Ubiquitin thioesterase FAF-X) (Ubiquitin-specific protease 9, X chromosome) (Ubiquitin-specific-processing protease FAF-X), USP9X, DFFRX FAM USP9

Species Reactivity

Human

Immunogen

Recombinant Antibody

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

11F2

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

USP9X is a large deubiquitinating enzyme that plays essential roles in regulating protein stability, cell signaling, and developmental pathways. As a member of the ubiquitin-specific protease family, USP9X removes ubiquitin from substrate proteins, thereby protecting them from proteasomal degradation. This enzymatic activity positions USP9X as a critical modulator of numerous cellular processes, including TGF-β signaling, Wnt pathway regulation, and chromosome segregation during mitosis. Dysregulation of USP9X has been implicated in various cancers and neurodevelopmental disorders, making it a compelling target for researchers investigating oncogenic mechanisms and therapeutic vulnerabilities.

This recombinant monoclonal antibody, clone 11F2, offers the reproducibility and consistency that demanding research applications require. Generated against a synthetic peptide derived from human USP9X, the antibody is produced using recombinant technology, ensuring sequence-defined specificity and eliminating the lot-to-lot variability often encountered with traditional hybridoma-derived antibodies. Affinity chromatography purification further enhances performance by reducing background signal.

Validation studies demonstrate robust detection across multiple experimental platforms. Western blot analysis confirms specific recognition of USP9X at the expected molecular weight of approximately 292 kDa across a diverse panel of human cell lines, including HeLa, U-251MG, MCF-7, K562, COLO-205, A549, and A431 cells. Immunohistochemistry staining has been successfully performed on paraffin-embedded human breast cancer and stomach tissue sections, while flow cytometry validation in HeLa cells confirms utility for intracellular protein detection.

This antibody serves researchers investigating ubiquitin biology, cancer cell signaling, and protein quality control mechanisms, providing a reliable tool for characterizing USP9X expression and localization across multiple experimental workflows.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Target Background

Function

Deubiquitinase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. May therefore play an important regulatory role at the level of protein turnover by preventing degradation of proteins through the removal of conjugated ubiquitin. Specifically hydrolyzes 'Lys-48'-, 'Lys-29'- and 'Lys-33'-linked polyubiquitins chains. Essential component of TGF-beta/BMP signaling cascade. Specifically deubiquitinates monoubiquitinated SMAD4, opposing the activity of E3 ubiquitin-protein ligase TRIM33. Deubiquitinates alkylation repair enzyme ALKBH3. OTUD4 recruits USP7 and USP9X to stabilize ALKBH3, thereby promoting the repair of alkylated DNA lesions. Regulates chromosome alignment and segregation in mitosis by regulating the localization of BIRC5/survivin to mitotic centromeres. Involved in axonal growth and neuronal cell migration. Regulates cellular clock function by enhancing the protein stability and transcriptional activity of the core circadian protein ARNTL/BMAL1 via its deubiquitinating activity.

Gene References into Functions

USP9X is overexpressed in breast carcinomas, and its level of expression is correlated with that of CEP131. USP9X is an important regulator of centrosome biogenesis and USP9X/CEP131 plays a critical role in breast carcinogenesis. PMID: 28361952
High USP9X expression is associated with Pancreatic Ductal Adenocarcinoma. PMID: 29248719
USP9X is a deubiquitylase of the Hippo pathway kinase LATS2 and that the Hippo pathway functions as a downstream signaling cascade that mediates USP9X's tumor-suppressive activity. PMID: 29183995
Ets-1 is induced by BRAF or MEK kinase inhibition, resulting in increased NRAS expression, which could be blocked by inactivation of Usp9x. PMID: 28198367
LNC473 could recruit deubiquitinase USP9X to inhibit the ubiquitination level of survivin and then increase survivin expression. PMID: 29605299
Study shows that USP9X deubiquitylating enzyme maintains RAPTOR protein levels, mTORC1 signalling and proliferation in neural progenitors. USP9X is the first deubiquitylating enzyme shown to stabilize RAPTOR. PMID: 28341829
Deubiquitylating enzyme USP9X was overexpressed in gastric cancer, suggesting a potential implication as an oncogene, and was significantly associated with a poorer survival. PMID: 28274596
Study demonstrated that USP9X acts as a tumor metastasis supporter in pancreatic ductal adenocarcinoma (PDAC), and its downregulation inhibited the migration and invasion of PDAC cells, while high expression of USP9X inhibited the apoptosis of cancer cells. Therefore, USP9X functioned as an oncogene in PDAC cells and is closely related with the expression of Snail, Twist and survivin. PMID: 29130109
knockdown of USP9X was shown to confer resistance to apoptosis following pediatric T-cell acute lymphoblastic leukemia relevant chemotherapy drug treatment in Jurkat leukemia cells PMID: 27602765
USP9X stabilizes beta-catenin and activates Wnt/beta-catenin signal pathway to promote glioma cell proliferation and survival. PMID: 27783990
Loss of USP9X expression is associated with pancreatic cancer. PMID: 28720576
Frame shift mutation in USP9X and deletion of the 5'UTR of the USP9X identified in two females with intellectual disability syndrome. PMID: 28377321
High USP9X expression is associated with basal-like breast cancer. PMID: 27593927
The authors find that primary human aggressive B-cell lymphoma samples exhibit high USP9X expression that correlate with XIAP overexpression. PMID: 27317434
USP9x-SMAD4 Interaction is associated with Breast Cancer Metastasis. PMID: 28115363
Data suggest that USP9X as an integral component of centrosome where it functions to stabilize PCM1 and CEP55 and to promote centrosome biogenesis; N-terminal domain of USP9X appears to be responsible for physical association of USP9X with PCM1 and CEP55. (USP9X = ubiquitin-specific protease 9X; PCM1 = pericentriolar material 1 protein; CEP55 = 55kDa centrosomal protein) PMID: 28620049
USP9X recruited to the centrosome by NPHP5 protects NPHP5 from ubiquitination, thus favouring cilia assembly. PMID: 28498859
BAG3 was found to positively regulate Mcl-1 levels by binding to and inhibiting USP9X. Our data show that BAG3 and Mcl-1 are key mediators of resistance to chemotherapy in ovarian cancer PMID: 27120977
found that USP9X regulated the expression and stability of CLASPIN in an S-phase-specific manner. USP9X depletion profoundly impairs the progression of DNA replication forks, causing unscheduled termination events with a frequency similar to CLASPIN depletion, resulting in excessive endogenous DNA damage PMID: 26921344
USP9X possibly promotes head and neck cancer cell proliferation through the mTOR pathway. PMID: 27374971
biological relevance of the ICP0- USP9X complex in HSV-1 infection PMID: 26596467
identify USP9X as a novel regulator of EGFR endocytosis PMID: 26748853
study reports for the first time that USP9X is a deubiquitinase of Angiomotin-like 2 (AMOTL2) and that AMOTL2 mono-ubiquitination is required for YAP inhibition PMID: 26598551
USP9x interacted with and stabilized beta-catenin through deubiquitination to mediate transcription of the decoy receptors in breast cancer cells PMID: 26717875
We examined the role of USP9X in the primary cilium of affected females with ciliopathy syndromes and found that endogenous USP9X localizes along the length of the ciliary axoneme, so its loss of function could disrupt cilium-regulated processes. PMID: 26833328
These data support the use of MCL1 expression as a predictive biomarker for USP9X inhibitors in non-small cell lung cancer therapy PMID: 25692226
The mammalian PRICKLE-interactome was defined, identifying prickle-interacting proteins that localize to synapses and a novel interacting partner, USP9X, a substrate-specific de-ubiquitinase. PRICKLE and USP9X interact through their carboxy-termini; and USP9X de-ubiquitinates PRICKLE, protecting it from proteasomal degradation. PMID: 25763846
the novel compound EOAI3402143 dose-dependently inhibited Usp9x and Usp24 activity, increased tumor cell apoptosis, and fully blocked or regressed myeloma tumors in mice. PMID: 25814533
Noxa upregulation reduces the availability of Usp9x to Mcl-1, thereby promoting its ubiquitination and degradation, leading to the apoptosis of neoplastic cells. PMID: 24991768
Although USP9X may function as a tumor-suppressor during the establishment of PDAC, data presented here argue that USP9X promotes cell growth in advanced PDAC cells when PDAC is typically diagnosed. PMID: 24841553
Our analysis of clinical HCC samples verifies that miR-26b also targets USP9X expression to inhibit the EMT of hepatocytes PMID: 24890815
USP9X downregulation renders breast cancer cells resistant to tamoxifen. PMID: 25028367
USP9X mutations may have a role in X-linked intellectual disability and disrupt neuronal cell migration and growth PMID: 24607389
a large set of SOX2-associated proteins in DAOY medulloblastoma cells PMID: 23667531
Mcl-1, Bcl-xL and USP9X overexpression are tumor survival mechanisms protective against chemotherapy. PMID: 23171055
The resulting USP9X-deficient cancer cells exhibited increased activation of apoptotic pathways. PMID: 22895071
USP9X is an important regulatory protein of SMURF1. PMID: 23184937
Usp9x plays an important role in stabilizing SMN and the SMN complex, likely via antagonizing Ub-dependent SMN degradation. PMID: 23112048
the deubiquitinase USP9X as a novel mTORC1 and -2 binding partner that negatively regulates mTOR activity and skeletal muscle differentiation. PMID: 22544753
loss of Usp9x enhances transformation and protects pancreatic cancer cells from anoikis PMID: 22699621
Our results suggest that ubiquitin-specific protease 9X (USP9X) is by far the most active deubiquitinase acting on Ub-PEX5, both in female rat liver and HeLa cells. PMID: 22371489
alpha-Synuclein fate is determined by USP9X-regulated monoubiquitination PMID: 22065755
Zymophagy, a novel selective autophagy pathway mediated by VMP1-USP9x-p62, prevents pancreatic cell death. PMID: 21173155
Ubiquitin-cycle inhibition represents a novel and effective approach to blocking Bcr-Abl kinase signaling and reducing Mcl-1 levels to engage chronic myelogenous leukemia cell apoptosis. PMID: 21248063
the deubiquitinase USP9X stabilizes MCL1 and thereby promotes cell survival; deubiquitinases may stabilize labile oncoproteins in human malignancies PMID: 20023629
FAM associates with E-cadherin and beta-catenin during trafficking to the plasma membrane PMID: 14742711
identified the ubiquitin-protease FAM/USP9X as a binding partner of Itch. The association between Itch and FAM/USP9X was confirmed in vitro by glutathione S-transferase pulldown and in vivo through coimmunoprecipation PMID: 17038327
NUAK1 and MARK4 are substrates of USP9X PMID: 18254724
Smad4 is monoubiquitinated in lysine 519 in vivo, a modification that inhibits Smad4 by impeding association with phospho-Smad2; FAM reverts this negative modification, re-empowering Smad4 function. PMID: 19135894

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Involvement in disease

Mental retardation, X-linked 99 (MRX99)

Subcellular Location

Cytoplasm. Cell projection, growth cone.

Protein Families

Peptidase C19 family

Tissue Specificity

Widely expressed in embryonic and adult tissues.

Database Links

HGNC: 12632

OMIM: 300072

KEGG: hsa:8239

STRING: 9606.ENSP00000316357

UniGene: Hs.77578

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Code	CSB-RA183712A0HU
Size	US$210
	Order now
Image	Western Blot Positive WB detected in: Hela whole cell lysate(30µg), U-251MG whole cell lysate(30µg), MCF-7 whole cell lysate(30µg), K562 whole cell lysate(30µg), COLO-205 whole cell lysate(30µg), A549 whole cell lysate(30µg), A431 whole cell lysate(30µg) All lanes: USP9X antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 290 kDa Observed band size: 292 kDa Exposure time：30s IHC image of CSB-RA183712A0HU diluted at 1:100 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-RA183712A0HU diluted at 1:100 and staining in paraffin-embedded human stomach tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Overlay Peak curve showing Hela cells stained with CSB-RA183712A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.
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USP9X Recombinant Monoclonal Antibody

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