- PA5-52147 - Invitrogen Antibodies SEC31A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of SEC31A was achieved by transfecting Hep G2 with SEC31A specific siRNAs (Silencer® select Product # s22550, s22551). Western blot analysis (Fig. a) was performed using membrane enriched cell extracts from the SEC31A knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with SEC31A Polyclonal Antibody (Product # PA5-52147, 0.4 µg/ml) 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 SEC31A.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-SEC31A Rabbit Polyclonal Antibody (Product # PA5-52147) and a 110 kDa band corresponding to SEC31A was observed across cell lines tested. Membrane enriched cell extracts (30 µg lysate) of A-431 (Lane 1), A549 (Lane 2), Hep G2 (Lane 3), K-562 (Lane 4), and SH-SY5Y (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were probed with the primary antibody (0.4 µg/ml), Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent staining of SEC31A in human cell line U-251 MG shows positivity in cytoplasm & vesicles. Samples were probed using a SEC31A Polyclonal Antibody (Product # PA5-52147).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of SEC31A was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with SEC31A Polyclonal Antibody (Product # PA5-52147) at 2 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then 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 staining in membrane and cytoplasm. 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 staining of SEC31A in human endometrium using SEC31A Polyclonal Antibody (Product # PA5-52147) shows positivit in glandular cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of SEC31A in human cerebral cortex using SEC31A Polyclonal Antibody (Product # PA5-52147) shows positivity in neurons.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of SEC31A in human liver using SEC31A Polyclonal Antibody (Product # PA5-52147) shows positivity in hepatocytes.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of SEC31A in human skeletal muscle using SEC31A Polyclonal Antibody (Product # PA5-52147) shows very weak positivity in myocytes.

PA5-52147