Oct4 Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb559071
• Category: Antibodies
• Description: Oct4 Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 2D5
• Tested applications: ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: Recombinant human Oct 4 protein, around 1-100aa
• Antibody Type: Recombinant Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:50-200, ICC/IF=1:50-200, IF=1:100-500
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 39 kDa
• Target: POU5F1
• UniProt ID: Q01860
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: MGC22487; Oct 3; Oct 4; Oct-3; Oct-4; OCT3; Oct4;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

ICC staining of Oct4 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (orb559071, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000 dilution. The nuclear counter stain is DAPI (blue).

ICC staining of Oct4 in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (orb559071, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemical analysis of paraffin-embedded human seminoma tissue tissue with Rabbit anti-Oct4 antibody at 1/4000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (orb559071) at 1/4000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Oct4 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (orb559071, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Western blot analysis of Oct4 on different lysates with Rabbit anti-Oct4 antibody (orb559071) at 1/500 dilution. Lane 1: HES cell lysate, Lane 2: NCCIT cell lysate, Lysates/proteins at 10 µg/Lane. Predicted band size: 39 kDa, Observed band size: 45 kDa, Exposure time: 1 minute, 10% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (orb559071) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300000 dilution was used for 1 hour at room temperature.

Western blot analysis of Oct4 on F9 cell lysates with Rabbit anti-Oct4 antibody (orb559071) at 1/1000 dilution. Lysates/proteins at 10 µg/Lane. Predicted band size: 39 kDa, Observed band size: 45 kDa, Exposure time: 2 minutes, 12% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (orb559071) at 1/1000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300000 dilution was used for 1 hour at room temperature.