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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
- 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
- 0.49 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 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.
- 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
- 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.
- 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 (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)
- 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
- 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.
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.
