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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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- Product number
- MA5-15585 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- USP7 Monoclonal Antibody (5F11)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15585 targets HAUSP in WB applications and shows reactivity with Human samples. The MA5-15585 immunogen is purified recombinant fragment of human HAUSP expressed in E. Coli.
- Antibody clone number
- 5F11
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HAUSP using HAUSP monoclonal antibody (Product # MA5-15585) in MCF-7 (1) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of USP7 was achieved by transfecting A549 with USP7 specific siRNAs (Silencer® select Product # S15441, S15440). Western blot analysis (Fig. a) was performed using Whole cell extracts from the USP7 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with USP7 Monoclonal Antibody (5F11) (Product # MA5-15585, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 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 USP7.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-USP7 Monoclonal Antibody (5F11)(Product # MA5-15585) and a 128kDa band corresponding to USP7 was observed across cell line and tissue tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), HT-29 (Lane 2), HeLa (Lane 3), HCT 116 (Lane 4), MCF7 (Lane 5) and tissue extract of Rat Kidney (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
