Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Rat, Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Primate - Macaca mulatta (Rhesus Macaque), Rabbit, Xenograft

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP), Bioassay, Co-Immunoprecipitation, Differentiation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Oct-3/4 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Oct‑3/4
Met1-Asn265 (Met262Leu)
Accession # Q01860

Specificity

Detects human Oct-3/4 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse Oct‑3/4 Antibody

Detection of Human and Mouse Oct-3/4 antibody by Western Blot.

Detection of Human and Mouse Oct‑3/4 by Western Blot.

Western blot shows lysates of BG01V human embryonic stem cells and D3 mouse embryonic stem cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Oct-3/4 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Alkaline Phosphatase and Oct‑3/4 antibody in BG01V Human Stem Cells by Immunocytochemistry (ICC).

Alkaline Phosphatase and Oct‑3/4 in BG01V Human Stem Cells.

Alkaline phosphatase (ALPL) and Oct-3/4 were detected in human BG01V embryonic stem cells using 10 µg/mL Human/Mouse/Rat ALPL Monoclonal Antibody (Catalog # MAB1448) and 10 µg/mL Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759). Cells were incubated with primary antibodies for 3 hours at room temperature. Cells were stained for ALPL using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (pseudo-stained green; Catalog # NL007), and stained for Oct-3/4 using the NorthernLights 637-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL002). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Oct-3/4-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of Oct‑3/4-regulated Genes by Chromatin Immunoprecipitation.

BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Oct-3/4/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thenanogpromoter was detected by standard PCR.

Detection of Human Oct-3/4 antibody by Simple WesternTM.

Detection of Human Oct‑3/4 by Simple WesternTM.

Simple Western lane view shows lysates of BG01V human embryonic stem cells and NTera-2 human testicular embryonic carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Oct-3/4 at approximately 50 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human Oct-3/4 by Western Blot

Detection of Human Oct-3/4 by Western Blot

Knockdown OGT inhibits proliferation and tumorsphere formation of hepatoma cell through reducing eIF4E expression. A, Huh7 and PLC/PRF/5 cells were infected with control shRNA, OGT shRNA2 alone, or with wild‐type eIF4E lentivirus. The cell lysates were harvested for western blotting analysis using indicated antibodies. beta ‐actin expression was served as a loading control. B, Cell proliferation of Huh7 and PLC/PRF/5 cells infected with lentiviruses as in panel (A) were measured with CCK8 assay. (C‐H) Huh7 and PLC/PRF/5 cells infected with lentiviruses as in panel (A) were seeded into 96‐well plates. After 12 d, tumorsphere were counted and quantified. Representative images of sphere (scale bars, 100 μm) were shown (C, F). The diameter of sphere (D, G) and number of sphere (E, H) were count. Data represent mean ± SD of at least three independent experiments. The two‐tailed Student's t tests were used. **P < 0.01. I, Huh7 cells expressing either OGT shRNA2 alone or with wild‐type eIF4E lentivirus were incubated with PE‐labelled anti‐AC133 antibody. The percentages of CD133+ cells in graphs were analysed by flow cytometry. Black line, control IgG staining; red line, CD133 staining. J, Cell lysates were examined by western blotting with indicated antibodies. The right panel showcases relative protein amounts of different groups. Error bars represent ±SD of triplicate experiments. **P < 0.01; n.s, no significance. K, Huh7 cells were collected and subjected to immunoprecipitation with antibody against eIF4E or normal mouse IgG. Total RNAs were purified from immunocomplexes and subjected to RT‐PCR to measure Sox2, OCT4, and KLF4 mRNAs associated with eIF4E Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30677218), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Oct-3/4 by Immunocytochemistry/Immunofluorescence

Detection of Human Oct-3/4 by Immunocytochemistry/Immunofluorescence

Neural induction of hPSCs. (a) Both hPSC culture and neural differentiation were performed on LN521 substrate. Cortical neurons were induced with dual SMAD inhibition, expanded in the presence of FGF2, and matured with support by a selection of neurotrophic factors. Neural progenitor cells (NPCs) could be cryopreserved at day 21 and plated at day 32 for final experiments, including microelectrode array (MEA) measurements. (b) Three different hPSC lines (one hESC line, 08/023 and two hiPSC lines, 10212.EURCCs and IMR90-4) were characterized for their efficiency in producing neuroectodermal cells in response to 12-day neural induction by dual SMAD inhibition. Cells were stained for pluripotency marker Oct4 at the pluripotent stage and after 12 days of neural induction. The presence of early neuroectodermal markers was evaluated with Sox2, FoxG1 and Pax6 staining. (c) After 21 days of differentiation, the culture contained vimentin- and Pax6-positive NPCs that could be cryopreserved. Additionally, the first Tbr2- and MAP2-positive neurons were detected at this time point. (d) The percentages of Pax6-positive cells were quantified at four time points of differentiation (mean ± s.e.m., n = 5–14, data derived from 1–3 independent differentiations). Statistical analysis was performed with the Mann-Whitney U test to compare differences between day 12 and day 46 within each hPSC line, and significant p-values are presented in the image. The scale bar is 50 µm in all images. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31748598), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse Oct‑3/4 Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/106 cells
Sample: BG01V human embryonic stem cell chromatin, for detection see our available Human Nanog Primer Pair (Catalog # RDP-320-025)

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed BG01V human embryonic stem cells

Immunoprecipitation

3 µg/106 cells
Sample: NTera‑2 human testicular embryonic carcinoma cell line, see our available Western blot detection antibodies

Simple Western

20 µg/mL
Sample: BG01V human embryonic stem cells and NTera‑2 human testicular embryonic carcinoma cell line

Western Blot

0.5 µg/mL
Sample: BG01V human embryonic stem cells and D3 mouse embryonic stem cell line

Reviewed Applications

Read 4 reviews rated 4.5 using AF1759 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Oct-3/4

Oct-3/4, a member of POU transcription factors, was identified as a DNA-binding protein that activates gene transcription via a cis-element containing an octamer motif (1). It is expressed in totipotent embryonic stem and germ cells (2, 3). A critical amount of Oct-3/4 expressed is required to sustain stem cell self-renewal and pluripotency (4). When embryonic stem cells are induced to differentiate, Oct-3/4 is downregulated and this downregulation of Oct-3/4 has proven to be essential for proper and divergent developmental program (5). The Oct-3/4 molecule is not only a master regulator of pluripotency that controls the lineage commitment but also is the most recognized marker used for the identification of totipotent embryonic stem cells.

References

  1. Scholer, H.R. et al. (1990) Nature 344:435.
  2. Scholer, H.R. et al. (1989) EMBO J. 8:2543.
  3. Rosner, M.H. et al. (1990) Nature 345:686.
  4. Niwa, H. et al. (2000) Nat. Genet. 24:372.
  5. Pesce, M. et al. (2001) Stem Cells 19:271.

Long Name

Octamer-binding Protein 3/4

Alternate Names

3-Oct, Oct4, Otf3g, Otf4, Pou5f1

Entrez Gene IDs

5460 (Human); 18999 (Mouse)

Gene Symbol

POU5F1

UniProt

Q01860

Additional Oct-3/4 Products

Product Documents for Human/Mouse Oct‑3/4 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse Oct‑3/4 Antibody

For research use only

Citations for Human/Mouse Oct‑3/4 Antibody

Customer Reviews for Human/Mouse Oct‑3/4 Antibody (4)

4.5 out of 5
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human/Mouse Oct-3/4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: BG01V human embryonic stem cells
    Species: Human
    Verified Customer | Posted 09/12/2025
  • Human/Mouse Oct-3/4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: human induced pluripotent stem cells
    Species: Human
    Verified Customer | Posted 10/24/2023
    Oct4 staining performed on iPSCs.
    Human/Mouse Oct‑3/4 Antibody AF1759
  • Human/Mouse Oct-3/4 Antibody
    Name: Shuba Krishnan
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: iPS2 human induced pluripotent stem cells
    Species: Human
    Verified Customer | Posted 10/18/2018
    Cells: human induced pluripotent stem cells Primary antibody: Goat anti-human OCT3/4 (1:200), O/N incubation at 4 degrees Secondary antibody: anti-goat AF568
    Human/Mouse Oct‑3/4 Antibody AF1759
  • Human/Mouse Oct-3/4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 11/23/2017
    Human/Mouse Oct‑3/4 Antibody AF1759

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