Invitrogen

USP9X Monoclonal Antibody (OTI2G7)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

View all (22) USP9X antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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USP9X Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Probable ubiquitin carboxyl-terminal hydrolase FAF-X was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.01% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with USP9X Monoclonal Antibody (OTI2G7) (Product # MA5-26367) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification.

USP9X Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human ovary tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a USP9X monoclonal antibody (Product # MA5-26367).

USP9X Antibody in Western Blot (WB) — Western Blot was performed using Anti-USP9X Monoclonal Antibody (OTI2G7) (Product # MA5-26367) and a 260 kDa band corresponding to Probable ubiquitin carboxyl-terminal hydrolase FAF-X was observed across across the cell lines tested. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), Hep G2 (Lane 2), MCF7 (Lane 3), PC-3 (Lane 4) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

USP9X Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HEK-293 cells were transfected with Probable ubiquitin carboxyl-terminal hydrolase FAF-X siRNA and decrease in signal intensity was observed in Western Blot application using Anti-USP9X Monoclonal Antibody (OTI2G7) (Product # MA5-26367). {KD}

Product Details

MA5-26367

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:2,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:150	-
Immunocytochemistry (ICC/IF)	1:100	-

Product Specifications
Species Reactivity	Dog, Human, Mouse, Non-human primate, Rat
Host/Isotype	Mouse / IgG2a
Class	Monoclonal
Type	Antibody
Clone	OTI2G7
Immunogen	Human recombinant protein fragment corresponding to amino acids 2246-2570 of USP9X produced in E.coli
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Affinity Chromatography
Storage buffer	PBS, pH 7.3, with 1% BSA, 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2723770

Target Information

USP9X is an ubiquitin-specific protease involved in the removal and recycling of ubiquitin from proteins degraded in proteasomes. As a deubiquitinating enzyme, USP9x has a critical function in tight junction biogenesis by controlling the levels of the exchange factor for Arf6 (EFA6), a protein shown to facilitate tight junction formation during a narrow temporal window preceding the establishment of cell polarity. USP9X is also located on the X-chromosome but escapes X-inactivation, and defects in USP9X may cause gonadal degeneration in Turner syndrome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Deubiquitinating enzyme FAF-X; Drosophila fat facets related, X-linked; Fat facets homolog; Fat facets in mammals; fat facets protein related, X-linked; Fat facets protein-related, X-linked; fats facets protein related, X; hFAM; Probable ubiquitin carboxyl-terminal hydrolase FAF-X; Ubiquitin carboxyl-terminal hydrolase FAM; ubiquitin specific peptidase 9, X chromosome; ubiquitin specific protease 9 SSSRF- isoform; ubiquitin specific protease 9, X chromosome; ubiquitin specific protease 9, X chromosome (fat facets-like Drosophila); Ubiquitin thioesterase FAF-X; ubiquitin thiolesterase FAF-X; ubiquitin-specific processing protease FAF-X; Ubiquitin-specific protease 9, X chromosome; Ubiquitin-specific-processing protease FAF-X

Gene Aliases: 5730589N07Rik; AA407302; AA407699; AL022658; AL022749; DFFRX; FAF; FAF-X; Fafl; FAM; MRX99; MRXS99F; RGD1560056; USP9; USP9X

UniProt ID: (Human) Q93008, (Mouse) P70398

Entrez Gene ID: (Dog) 480885, (Human) 8239, (Mouse) 22284, (Rat) 363445

Molecular Function: cysteine protease