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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunoprecipitation [2]
- Immunohistochemistry [4]
- Other assay [2]
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- Product number
- PA5-28209 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- OCT1 (POU2F1) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, HeLa, HepG2. Predicted reactivity: Mouse (89%), Rat (94%), Xenopus laevis (81%), Pig (97%), Chicken (94%), Chimpanzee (99%), Bovine (95%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 (Heavy Chain) 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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)
- Main image
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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)
- Main image
- 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.
- 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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)
- Main image
- 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.
- 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.
