PA5-20087

antibody from Invitrogen Antibodies

Targeting: IRF3

Antibody data

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Validation data

IRF3 protein structure - PA5-20087 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of U937 whole cell lysate using a IRF3 polyclonal antibody (Product # PA5-20087) at (A) 1, (B) 2, and (C) 4 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of IRF3 in (A) human kidney and (B) rat kidney lysate with IRF3 Polyclonal Antibody (Product # PA5-20087) at 1 µg/mL..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of IRF3 was achieved by transfecting A-431 cells with IRF3 specific siRNAs (Silencer® select Product # s7509, s7507). Western blot analysis (Fig. a) was performed using membrane extracts from the IRF3 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with IRF3 Polyclonal Antibody (Product # PA5-20087, 1 µg/mL) 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 IRF3.

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 K-562 (Lane 1), THP-1 (Lane 2), HL-60 (Lane 3), U-937 (Lane 4), A549 (Lane 5), A-431 (Lane 6), MCF7 (Lane 7) and HT-29 (Lane 8). The blot was probed with Anti-IRF3 Polyclonal Antibody (Product # PA5-20087, 4 µg/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). 50kDa and 33kDa bands corresponding to IRF3 isoforms were observed across all the cell lines tested. An uncharacterized band was observed at ~80kDa in certain cell lines.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Ramos cells using a IRF3 polyclonal antibody (Product # PA5-20087) at a 2 µg/mL dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry of IRF3 in Ramos cells with IRF3 Polyclonal Antibody (Product # PA5-20087) at 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence of IRF3 in Ramos cells with IRF3 Polyclonal Antibody (Product # PA5-20087) at 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of IRF3 was performed using 70% confluent log phase MCF7 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 IRF3 Polyclonal Antibody (Product # PA5-20087) at 1:100 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) 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, 1:300). Panel d represents the merged image showing nucleus and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.