- PA5-27856 - Invitrogen Antibodies RRM2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RRM2 using 30 µg of H1299 lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a RRM2 polyclonal antibody (Product # PA5-27856) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of RRM2 was achieved by transfecting A549 cells with RRM2 specific siRNAs (Silencer® select Product # s12360, s12361). Western blot analysis (Fig. a) was performed using whole cell extracts from the RRM2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and un-transfected cells (lane 1). The blots were probed with RRM2 Polyclonal Antibody (Product # PA5-27856, 1µg/ml) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometry 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 RRM2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), HeLa (Lane 2), K-562 (Lane 3), Jurkat (Lane 4) and Ramos (Lane 5). The blot was probed with Anti-RRM2 Polyclonal Antibody (Product # PA5-27856, 1µg/ml) 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 44 kDa band corresponding to RRM2 was observed across all the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of RRM2 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: RRM2 Polyclonal Antibody (Product # PA5-27856) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of RRM2 was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: RRM2 Polyclonal Antibody (Product # PA5-27856) diluted at 1:1000. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of RRM2 was performed using 70% confluent log phase A549 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 RRM2 Polyclonal Antibody (Product # PA5-27856) at 5 µg/mL 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant cytosolic 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: Immunohistochemistry (Paraffin) analysis of RRM2 was performed in paraffin-embedded rat intestine tissue using RRM2 Polyclonal Antibody (Product # PA5-27856) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of RRM2 was performed in paraffin-embedded mouse muscle tissue using RRM2 Polyclonal Antibody (Product # PA5-27856) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of RRM2 was performed in paraffin-embedded rat intestine tissue using RRM2 Polyclonal Antibody (Product # PA5-27856) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: RRM2 Polyclonal Antibody detects RRM2 protein at cytoplasm on mouse muscle by immunohistochemical analysis. Sample: Paraffin-embedded mouse muscle. RRM2 Polyclonal Antibody (Product # PA5-27856) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-27856