- PA5-31144 - Invitrogen Antibodies RAD51 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of RAD51 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a RAD51 Polyclonal Antibody (Product # PA5-31144) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using RAD51 Polyclonal Antibody (Product # PA5-31144). Sample (30 µg of whole cell lysate). Lane A: JurKat. 10% SDS PAGE. RAD51 Polyclonal Antibody (Product # PA5-31144) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of RAD51 was achieved by transfecting HeLa with RAD51 specific siRNAs (Silencer® select Product # s11734, s11735). Western blot analysis (Fig. a) was performed using whole cell extracts from the RAD51 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with RAD51 Polyclonal Antibody (Product # PA5-31144, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to RAD51.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of NIH/3T3 (Lane 1), HeLa (Lane 2), RAW 264.7 (Lane 3), Neuro-2A (Lane 4), Jurkat (Lane 5), Raji (Lane 6) and PANC-1 (Lane 7). The blot was probed with Anti-RAD51 Polyclonal Antibody (Product # PA5-31144, 1:1000 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). Band of ~40 kDa corresponding to RAD51 was observed in cell lines tested along with additional bands at ~78 kDa and 48 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of RAD51 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RAD51 Polyclonal Antibody (Product # PA5-31144) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing nuclear, cytoplasmic and perinuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: RAD51 Polyclonal Antibody detects Rad51 protein at cytosol and nucleus on human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. RAD51 Polyclonal Antibody (Product # PA5-31144) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: RAD51 Polyclonal Antibody detects Rad51 protein at cytosol on human ovarian carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human ovarian carcinoma. RAD51 Polyclonal Antibody (Product # PA5-31144) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: RAD51 Polyclonal Antibody immunoprecipitates Rad51 protein in IP experiments. IP samples: Jurkat whole cell extract. A. 40 µg Jurkat whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of Rad51 protein by 4 µg RAD51 Polyclonal Antibody (Product # PA5-31144). 5 % SDS-PAGE. The immunoprecipitated Rad51 protein was detected by RAD51 Polyclonal Antibody (Product # PA5-31144) diluted at 1:500.

PA5-31144