Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- ELISA [1]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-31935 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TRIM25 Monoclonal Antibody (5B5B12)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 5B5B12
- 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 MCF-7 (1), MCF-7 (2), K562 (3), A549 (4), and MOLT4 (5) cell lysate. Samples were incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:500-1:2000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRIM25 in human TRIM25 recombinant protein. Sample was incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:500-1:2000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TRIM25 in HEK293 (1) and TRIM25 cell lysate. Samples were incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:500-1:2000.
- 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 Monoclonal Antibody (5B5B12) (Product # MA5-31935, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 an uncharacterised (*) band at 59 kDa.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western Blot was performed using Anti-TRIM25 Monoclonal Antibody (5B5B12) (Product # MA5-31935) and a 70 kDa band corresponding to E3 ubiquitin/ISG15 ligase TRIM25 was observed across tested samples along with an uncharacterised (*) band at 59 kDa. Whole cell extracts (40 µg lysate) of SK-O-V3 (Lane 1), SK-BR-3 (Lane 2), HCT 116 (Lane 3), MCF7 (Lane 4), K-562 (Lane 5), NIH/3T3 (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, 1:20000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ELISA analysis of TRIM25 in Control Antigen (black line, 100 ng); Antigen (purple line, 10 ng); Antigen (blue line, 50 ng); Antigen (red line, 100 ng). Samples were incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:10,000.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of TRIM25 in Hela cells (green). Sample was incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:50-1:250 followed by DRAQ5 fluorescent DNA dye (blue), and Alexa Fluor- 555 phalloidin (red labeled actin filaments) .
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry analysis of TRIM25 in paraffin-embedded bladder cancer tissue. Sample was incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:200-1:1000 followed by DAB staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of TRIM25 in paraffin-embedded rectum cancer tissue. Sample was incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:200-1:1000 followed by DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry of TRIM25 in HeLa cells (green). Sample was incubated with TRIM25 monoclonal antibody (Product # MA5-31935) using a dilution of 1:200-1:400 followed by negative control (red).