Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [6]
- Immunocytochemistry [4]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-23035 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- USP9X Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- The target sequence has 95% sequence homology with rat and 80% sequence homology with zebrafish.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.2 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of USP9X using a polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of USP9X using a polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of USP9X in Caco-2 whole cell lysates. Sample was incubated in USP9X polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of USP9X in NIH/3T3 whole cell lysate. Sample was incubated in USP9X polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Probable ubiquitin carboxyl-terminal hydrolase FAF-X was achieved by transfecting HEK-293 with Probable ubiquitin carboxyl-terminal hydrolase FAF-X specific siRNAs (Silencer® select Product # s15742, s15743). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Probable ubiquitin carboxyl-terminal hydrolase FAF-X knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with USP9X Polyclonal Antibody (Product # PA5-23035, 1:1000 dilution ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution). Densitometric analysis of this western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Probable ubiquitin carboxyl-terminal hydrolase FAF-X.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-USP9X Polyclonal Antibody (Product # PA5-23035) 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of USP9X using a polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of USP9X in NTERA-2 cells. Samples were incubated in USP9X polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of USP9X in NTERA-2 cells. Samples were incubated in USP9X polyclonal antibody (Product # PA5-23035).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with USP9X Polyclonal Antibody (Product # PA5-23035) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034), (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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of USP9X in brain tissue. Samples were incubated in USP9X polyclonal antibody (Product # PA5-23035).