Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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Validation data
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- Product number
- PA5-32472 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Ku80 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 1mM EDTA buffer, pH 8.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is placenta tissue.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 500 µL
- 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
- Knockdown of Ku80 was achieved by transfecting HeLa with Ku80 specific siRNAs (Silencer® select Product # s14952, s14953). Western blot analysis (Fig. a) was performed using whole cell extracts from the Ku80 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Ku80 Polyclonal Antibody (Product # PA5-32472, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 Ku80.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Ku80 Polyclonal Antibody (Product # PA5-32472) and a 80 kDa band corresponding to Ku80 was observed in the cell lines tested. Modified whole cell extracts (1%SDS) (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), K-562 (Lane 3), Jurkat (Lane 4), COS-7 (Lane 5), and A549 (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).