Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [6]
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Validation data
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- Product number
- PA5-75252 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TRIM25 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRIM25 in Lane 1: HEK293T whole cell lysate (40 µg), Lane 2: MCF-7 whole cell lysate (40 µg). Samples were incubated with TRIM25 polyclonal antibody (Product # PA5-75252) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRIM25 in Lane 1: HEK293T whole cell lysate (40 µg), Lane 2: MCF-7 whole cell lysate (40 µg). Samples were incubated with TRIM25 polyclonal antibody (Product # PA5-75252) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRIM25 in Lane 1: HEK293T whole cell lysate (40 µg), Lane 2: MCF-7 whole cell lysate (40 µg). Samples were incubated with TRIM25 polyclonal antibody (Product # PA5-75252) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRIM25 in Lane 1: H1792 whole cell lysate (10 µg), Lane 2: HepG2 whole cell lysate (10 µg), Lane 3: MCF-7 whole cell lysate (10 µg), Lane 4: L02 whole cell lysate (10 µg). Samples were incubated with TRIM25 polyclonal antibody (Product # PA5-75252) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of E3 ubiquitin/ISG15 ligase TRIM25 was achieved by transfecting SK-BR-3 with E3 ubiquitin/ISG15 ligase TRIM25 specific siRNAs (Silencer® select Product # S15206, S15204). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the E3 ubiquitin/ISG15 ligase TRIM25 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TRIM25 Polyclonal Antibody (Product # PA5-75252, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 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 E3 ubiquitin/ISG15 ligase TRIM25 along with uncharacterised (*) bands at 50 and 15 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-TRIM25 Polyclonal Antibody (Product # PA5-75252) and a 70 kDa band corresponding to E3 ubiquitin/ISG15 ligase TRIM25 was observed across along with an uncharacterised (*) band at ~50 kDa. Whole cell extracts (40 µg lysate) of SK-O-V3 (Lane 1), PANC-1 (Lane 2), K-562 (Lane 3), SK-BR-3 (Lane 4), MCF7 (Lane 5), HCT 116 (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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).