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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
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- Product number
- PA1-41294 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DNA Ligase I Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Suggested positive control: Hela whole cell extract, HeLa cell lysate.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 200 µL
- Concentration
- 0.5 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 LIG1 in cell lysate from HeLa cells using a LIG1 polyclonal antibody (Product # PA1-41294) at 1:1000 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DNA Ligase I in HeLa cell lysate. Sample was incubated in DNA Ligase I polyclonal antibody (Product # PA1-41294).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DNA ligase 1 was achieved by transfecting HeLa with DNA ligase 1 specific siRNAs (Silencer® select Product # s8174, s8173). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the DNA ligase 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DNA Ligase I Polyclonal Antibody (Product # PA1-41294, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 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 DNA ligase 1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DNA Ligase I Polyclonal Antibody (Product # PA1-41294) and a 130kDa band corresponding to DNA ligase 1 was observed across cell lines tested. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), Raji (Lane 2), A-431 (Lane 3), A549 (Lane 4), HT-29 (Lane 5), MIA PaCa-2 (Lane 6), MCF7 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
