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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [7]
- Immunohistochemistry [1]
- Other assay [2]
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- Product number
- PA5-31554 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RAD50 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLa, NT2D1, Neuro2A, GL261, PC-12.
- Concentration
- 0.78 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Rad50 in HeLa whole cell and nuclear extracts (30 µg). Samples were separated by 5% SDS-PAGE and the membrane was probed with Rad50, N-term Polyclonal antibody (Product # PA5-31554) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on nuclear enriched cell extract (30 µg lysate) of HeLa (Lane 1), SH-SY5Y (Lane 2), U-87MG (Lane 3), Neuro2A (Lane 4), NTERA-2 (Lane 5), NIH/3T3 (Lane 6) and PANC-1 (Lane 7). The blot was probed with Anti-RAD50 Polyclonal Antibody (Product # PA5-31554, 1:5000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 153 kDa band corresponding to RAD50 was detected in cell lines tested with a non-specific band at ~34kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of RAD50 was performed by separating 30 µg of whole cell extract by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a RAD50 Polyclonal Antibody (Product # PA5-31554) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using RAD50 Polyclonal Antibody (Product # PA5-31554). Sample (30 µg of whole cell lysate). Lane A: NT2D1. 5% SDS PAGE. RAD50 Polyclonal Antibody (Product # PA5-31554) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using RAD50 Polyclonal Antibody (Product # PA5-31554). HeLa whole cell and nuclear extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with RAD50 Polyclonal Antibody (Product # PA5-31554) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- RAD50 Polyclonal Antibody detects RAD50 protein by western blot analysis. A. 30 µg Neuro2A whole lysate/extract. B. 30 µg GL261 whole cell lysate/extract.5% SDS-PAGE. RAD50 Polyclonal Antibody (Product # PA5-31554) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of RAD50 was achieved by transfecting U-87 MG cells with RAD50 specific siRNAs (Silencer® select Product # s791, s792). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the RAD50 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with RAD50 Polyclonal Antibody (Product # PA5-31554, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 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 RAD50.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Rad50 (Product # PA5-31554) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- RAD50 Polyclonal Antibody immunoprecipitates Rad50 protein in IP experiments. IP samples: NT2D1 whole cell extract. A. 50 µg NT2D1 whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of Rad50 protein by 4 µg RAD50 Polyclonal Antibody (Product # PA5-31554). 5 % SDS-PAGE. The immunoprecipitated Rad50 protein was detected by RAD50 Polyclonal Antibody (Product # PA5-31554) diluted at 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- RAD50 Polyclonal Antibody immunoprecipitates Rad50 protein in IP experiments. IP samples: NT2D1 whole cell extract. A. 50 µg NT2D1 whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of Rad50 protein by 4 µg RAD50 Polyclonal Antibody (Product # PA5-31554). 5 % SDS-PAGE. The immunoprecipitated Rad50 protein was detected by RAD50 Polyclonal Antibody (Product # PA5-31554) diluted at 1:500.
