Oct3 4 antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Oct3/4 antibody is a highly specific and sensitive tool used in research laboratories for the detection and analysis of the transcription factor Oct3/4, which is a key regulator of pluripotency in stem cells. This antibody recognizes the Oct3/4 protein and can be used in various experimental techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of Oct3/4 in different cell types and tissues.

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Lab products found in correlation

Vector red alkaline phosphatase staining kit, by Vector Laboratories (1 mentions) Iplabs software, by BD (1 mentions) Nanog antibody, by Abcam (1 mentions) Gelatin coated chamber slides, by Thermo Fisher Scientific (1 mentions) Anti mouse igg alexa 488 conjugate antibody, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Wuhan Servicebio Technology (1 mentions)

Spelling variants of this product

OCT3/4 (94 citations) Anti-Oct3/4 antibody (3 citations)

4 protocols using oct3 4 antibody

Pluripotency Marker Staining Protocol

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Vector Red alkaline phosphatase staining kit (Vector Labs) or Live AP (Life Technologies) at 1:100 dilution; Stainalive Tra-1-60 antibody (09-0068; Stemgent) at 1:200; and mouse monoclonal Oct3/4 antibody (sc-5279; Santa Cruz) 1:100 dilution with Alexa Fluor® 594 Donkey anti-mouse secondary at 1:300 dilution were used. Antibody incubations were performed at 4 °C. Cells were counter-stained with DAPI in PBS and imaged in PBS.

Schmitt C.E., Morales B.M., Schmitz E.M., Hawkins J.S., Lizama C.O., Zape J.P., Hsiao E.C, & Zovein A.C. (2017). Fluorescent tagged episomals for stoichiometric induced pluripotent stem cell reprogramming. Stem Cell Research & Therapy, 8, 132.

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Immunofluorescent Analysis of Pluripotency Markers

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Cells and blastocysts were plated on 0.1% gelatin-coated chamber slides (Thermo Fisher Scientific), treated as indicated, and then fixed with 3.7% formaldehyde diluted in PBS for 30 minutes. The slides were permeabilized with 0.1% Triton X-100 diluted in PBS, blocked with 10% BSA diluted in PBS, and incubated with an Oct3/4 antibody (Santa Cruz Biotechnology) and a Nanog antibody (Abcam) diluted in 5% BSA in PBS at a 1:200 dilution, for 2 hours. The slides were washed with PBS, and then were incubated with an anti-mouse IgG-Alexa 488 conjugate antibody (Thermo Fisher Scientific) and an anti-rabbit IgG-Alexa 548 conjugate antibody (Thermo Fisher Scientific) diluted in 5% BSA in PBS at a 1:400 dilution, for 1 hour. The slides were again washed with PBS and visualized by brightfield and fluorescent microscopy. All images were captured using IPLABS software (BD Biosciences) and processed using ImageJ software.

Hur Y.H., Feng S., Wilson K.F., Cerione R.A, & Antonyak M.A. (2020). Embryonic Stem Cell-Derived Extracellular Vesicles Maintain ESC Stemness by Activating FAK. Developmental cell, 56(3), 277-291.e6.

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Immunofluorescence Analysis of Pluripotency in IVF Embryos

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Embryos with 32 or more cells at 144 h post IVF were fixed in 4% paraformaldehyde and permeabilized with 1% Triton X-100 in PBS. Samples were blocked with 1% BSA and 10% normal donkey serum in DPBS and incubated overnight with goat anti-OCT4 primary antibody (1:300; OCT3/4 antibody, Santa Cruz Biotechnology, Santa Cruz, United States). After extensive washing, embryos were incubated for 1 h with anti-goat IgG Alexa 568 secondary antibody (1:500; Invitrogen, United States) and 20 min with 10 μg/mL Hoechst 33342. Samples were observed using an epi-fluorescence microscope (Revolve, Echo, San Diego, United States). Number of cells per embryo showing expression Hoechst and/or Alexa 568 fluorescence was recorded and means compared between treatments.

Camargo L.S., Owen J.R., Van Eenennaam A.L, & Ross P.J. (2020). Efficient One-Step Knockout by Electroporation of Ribonucleoproteins Into Zona-Intact Bovine Embryos. Frontiers in Genetics, 11, 570069.

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Immunostaining of Blastocyst OCT3/4

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After co-culture with oEVs for 72 h, blastocysts were fixed in 4% paraformaldehyde for 30 min. The fixed blastocysts were then permeabilized with 0.1% Triton X-100 (Servicebio, Wuhan, China) for 20 min, blocked in 3% BSA for 1 h, and then incubated with primary OCT3/4 antibody (Santa Cruz, Dallas, TX, USA) overnight at 4°C. Secondary antibody labeled with cyanine-3 was then used to mark the primary antibody. DNA was stained with 4 μg/ml Hoechst 33258 dye solution (Servicebio, Wuhan, China) for 10 min. The blastocysts were then mounted onto glass slides and observed under a confocal microscope (Nikon, Tokyo, Japan).

Li Y., Liu C., Guo N., Cai L., Wang M., Zhu L., Li F., Jin L, & Sui C. (2023). Extracellular vesicles from human Fallopian tubal fluid benefit embryo development in vitro. Human Reproduction Open, 2023(2), hoad006.

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