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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- MA1-23189 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DNA Ligase I Monoclonal Antibody (10H5)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Recommended positive controls: MCF7.
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 10H5
- Vial size
- 100 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 DNA Ligase I in Raji whole cell lysate using a DNA Ligase I monoclonal antibody (Product # MA1-23189).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DNA Ligase I Monoclonal Antibody (10H5) (Product # MA1-23189). Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with DNA Ligase I Monoclonal Antibody (10H5) (Product # MA1-23189) diluted at 1:2,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
- 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 Monoclonal Antibody (10H5) (Product # MA1-23189, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 Monoclonal Antibody (10H5) (Product # MA1-23189) and a 130kDa band corresponding to DNA ligase 1 was observed across cell lines and tissue tested. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), Raji (Lane 2), A-431 (Lane 3), A549 (Lane 4), MIA PaCa-2 (Lane 5), MCF7 (Lane 6), Mouse Testis (Lane 7) 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, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DNA Ligase I Monoclonal Antibody (10H5) detects DNA ligase I protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: DNA ligase I stained by DNA Ligase I Monoclonal Antibody (10H5) (Product # MA1-23189) diluted at 1:500. Red: phalloidin, a cytoskeleton marker, diluted at 1:200. Scale bar= 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of DNA Ligase 1 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with DNA Ligase I Monoclonal Antibody (10H5) (Product # MA1-23189) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DNA Ligase I Monoclonal Antibody (10H5) detects DNA ligase I protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. DNA ligase I stained by DNA Ligase I Monoclonal Antibody (10H5) (Product # MA1-23189) diluted at 1:100. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
