- PA5-28209 - Invitrogen Antibodies POU2F1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of OCT1 using 30 µg of A) 293T (B) A431 (C) HeLa and D) HepG2 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with an OCT1 (POU2F1) polyclonal antibody (Product # PA5-28209) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of OCT1 (POU2F1) was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209) at a dilution of 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of OCT1 was achieved by transfecting HeLa with OCT1 specific siRNAs (Silencer® select Product # s10848, s10847). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the OCT1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209, 0.5 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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 OCT1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti- OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209) and ~80 kDa band corresponding to OCT1 was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Ramos (Lane 2), HEK-293 (Lane 3) and HCT 116 (Lane 4) 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 blot was probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of OCT1 (POU2F1) was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: OCT1 (POU2F1) Polyclonal Antibody (Product # PA5 28209) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of OCT1 (POU2F1) was performed in paraffin-embedded rat brain tissue using OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded Hela xenograft, using OCT1 (Product # PA5-28209) antibody at 1:750 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: OCT1 (POU2F1) Polyclonal Antibody immunoprecipitates OCT1 protein in IP experiments. IP Sample: 293T whole cell lysate/extract A : Control with 2.5 µg of pre-immune rabbit IgG B : Immunoprecipitation of OCT1 by 2.5 µg of OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209) 7.5% SDS-PAGE The immunoprecipitated OCT1 protein was detected by OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209) diluted at 1:1,000. Anti-rabbit IgG (HRP) was used as a secondary reagent.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of OCT1 was performed in 293T whole cell extracts using 5 µg of OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-28209). Samples were transferred to a membrane and probed with OCT1 (POU2F1) Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PA5-28209