Recombinant Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13809(B)] to USP9x - N-terminal
- Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Human
Overview
-
Product name
Anti-USP9x antibody [EPR13809(B)] - N-terminal
See all USP9x primary antibodies -
Description
Rabbit monoclonal [EPR13809(B)] to USP9x - N-terminal -
Host species
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP MouseHumanWB MouseHuman -
Immunogen
Synthetic peptide within Human USP9x aa 1-100 (Cysteine residue). The exact sequence is proprietary.
Database link: Q93008 -
Positive control
- WB: HeLa, Hap1, T84, NIH3T3, Jurkat and HepG2 cell lysates. IHC-P: Human kidney and testis tissues. IP: HeLa and NIH3T3 cell lysates. Flow Cyt: Jurkat cells. ICC/IF: HeLa cells.
-
General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
-
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR13809(B) -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
KO cell pellets
-
Positive Controls
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab180191 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
---|---|
Flow Cyt |
Human
|
ICC/IF |
Human
|
IHC-P |
Human
|
IP |
Mouse
Human
|
WB |
Mouse
Human
|
All applications |
Rat
|
Application | Abreviews | Notes |
---|---|---|
IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
|
WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 292 kDa.
|
ICC/IF |
1/100 - 1/250.
|
|
IP |
1/20.
|
|
Flow Cyt |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
Notes |
---|
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
WB
1/1000 - 1/10000. Predicted molecular weight: 292 kDa. |
ICC/IF
1/100 - 1/250. |
IP
1/20. |
Flow Cyt
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
Target
-
Function
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. -
Tissue specificity
Widely expressed in embryonic and adult tissues. -
Sequence similarities
Belongs to the peptidase C19 family. -
Cellular localization
Cytoplasm. - Information by UniProt
-
Database links
- Entrez Gene: 8239 Human
- Entrez Gene: 22284 Mouse
- Entrez Gene: 363445 Rat
- Omim: 300072 Human
- SwissProt: Q93008 Human
- SwissProt: P70398 Mouse
- Unigene: 77578 Human
- Unigene: 242646 Mouse
-
Alternative names
- Deubiquitinating enzyme FAF X antibody
- Deubiquitinating enzyme FAF-X antibody
- DFFRX antibody
see all
Images
-
All lanes : Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : USP9X knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 292 kDa
Observed band size: 290 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab180191 observed at 290 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab180191 was shown to react with USP9x in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265665 (knockout cell lysate ab257790) was used. Wild-type HeLa and USP9X knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180191 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
Immunohistochemical analysis of paraffin embedded Human kidney tissue labeling USP9x with ab180191 at 1/50.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
ab180191 (purified) at 1/20 dilution immunoprecipitating ETS2 in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab180191 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab180191 in HeLa whole cell lysateFor western blotting, ab180191 at 1/500 dilution (0.23 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.
Blocking and diluting buffer: 5% NFDM /TBST.
-
Immunofluorescent analysis of HeLa cells labeling USP9x with ab180191 at 1/100 (green) and DAPI staining (blue).
-
Flow Cytometrical analysis of permeabilized Jurkat cells labeling USP9x with ab180191 at 1/10 (red) or a rabbit IgG negative (green).
-
ab180191 (purified) at 1/20 dilution immunoprecipitating ETS2 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg.
Lane 2 (+): ab180191 & NIH/3T3 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab180191 in NIH/3T3 whole cell lysateFor western blotting, ab180191 at 1/500 dilution (0.23 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.
Blocking and diluting buffer: 5% NFDM /TBST.
-
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 - 4: Merged signal (red and green). Green - ab180191 observed at 270 kDa. Red - loading control, ab18058, observed at 117 kDa.ab180191 was shown to specifically react with USP9x when USP9x knockout samples were used. Wild-type and USP9x knockout samples were subjected to SDS-PAGE. ab180191 and ab18058 (loading control to Vinculin) were 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 (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
-
All lanes : Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 292 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)
Immunohistochemical analysis of paraffin embedded Human testis tissue labeling USP9x with ab180191 at 1/50.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (1)
ab180191 has been referenced in 1 publication.
- Li H et al. VCP/p97 increases BMP signaling by accelerating ubiquitin ligase Smurf1 degradation. FASEB J 33:2928-2943 (2019). PubMed: 30335548