- PA5-13540 - Invitrogen Antibodies EEF1A1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis using an EEF1A1 polyclonal antibody (Product # PA5-13540) in Y79, T47D cell lysates (35 µg per lane).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis using 293T cell lysate. Resolved proteins were electrophoretically transferred to PVDF membrane and incubated sequentially with an EEF1A1 polyclonal antibody (Product # PA5-13540) (1:500, 4°C, overnight) followed by detection using an HRP-conjugated rabbit secondary antibody. After washing, the bound antibody complex was detected using an ECL chemiluminescence reagent.

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 SK-OV-3 (Lane 1), HCT 116 (Lane 2), PC-3 (Lane 3), THP-1 (Lane 4), K-562 (Lane 5), MCF7 (Lane 6), C2C12 (Lane 7) and HeLa (Lane 8). The blot was probed with Anti-EEF1A1 Polyclonal Antibody (Product # PA5-13540, 1:3000 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 50 kDa band corresponding to EEF1A1 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of EEF1A1 was achieved by transfecting MCF7 with EEF1A1 specific siRNAs (Silencer® select Product # s4477, s4478 ). Western blot analysis (Fig. a) was performed using whole cell extracts from the EEF1A1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with EEF1A1 Polyclonal Antibody (Product # PA5-13540, 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). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to EEF1A1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of EEF1A1 was performed using 70% confluent log phase MCF-7 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 EEF1A1 Rabbit Polyclonal Antibody(Product # PA5-13540) 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) 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, 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of formalin-fixed, paraffin-embedded human brain tissue using an EEF1A1 polyclonal antibody (Product # PA5-13540), followed by HRP-conjugated secondary antibody and DAB staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of NCI-H292 cells using an EEF1A1 polyclonal antibody (Product # PA5-13540) (bottom), compared to a negative control cell (top) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

PA5-13540