Antibodies, Proteins, Kits and Reagents for Life Science

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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

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	Human
ICC/IF	Human
IHC-P	Human
IP	Mouse Human
WB	Mouse Human

See all applications and species data

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
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.

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

Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

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.
Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

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 lysate

For 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.
Immunocytochemistry/ Immunofluorescence - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

Immunofluorescent analysis of HeLa cells labeling USP9x with ab180191 at 1/100 (green) and DAPI staining (blue).
Flow Cytometry - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

Flow Cytometrical analysis of permeabilized Jurkat cells labeling USP9x with ab180191 at 1/10 (red) or a rabbit IgG negative (green).
Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

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 lysate

For 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.
Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

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.
Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

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.
Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-USP9x antibody [EPR13809(B)] - N-terminal (ab180191)

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Images

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