Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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Validation data
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- Product number
- MA5-17260 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Coilin Monoclonal Antibody (1H10)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- A suggested positive control for this product is HeLa cell lysate.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 1H10
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Coilin using anti-Coilin monoclonal antibody (Product # MA5-17260) on (Lane 1) HeLa Cell lysate, (Lane 2) SK-N-MC Cell lysate, (Lane 3) K562 Cell lysate, (Lane 4) A431 Cell lysate, (Lane 5) HepG2 Cell lysate, (Lane 6) SK-BR3 Cell lysate and (Lane 7) Ramos Cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Coilin was achieved by transfecting HeLa with Coilin specific siRNAs (Silencer® select Product # s15663, s15662). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Coilin knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Coilin Monoclonal Antibody (1H10) (Product # MA5-17260, 0.5ug/ml ) 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 Coilin. An uncharacterized band at ~25 kDa was also observed.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Coilin Monoclonal Antibody (1H10) (Product # MA5-17260) and a 63kDa band corresponding to Coilin was observed across cell lines and tissue tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2), Hep G2 (Lane 3), A549 (Lane 4), A-431 (Lane 5), K-562 (Lane 6), Reh (Lane 7), U-2 OS (Lane 8), MCF7 (Lane 9), Rat Testis (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (0.5 ug/mL) 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). An uncharacterized band was observed at ~25 kDa.