PA5-31554

antibody from Invitrogen Antibodies

Targeting: RAD50 hRad50, RAD50-2

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-31554

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. Predicted reactivity: Mouse (92%), Rat (91%), Rabbit (92%), Bovine (93%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.78 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Immunohistochemistry

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.

Other assay

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.

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.