Human/Mouse Oct-3/4 Antibody

(17 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human Oct-3/4 in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Oct‑3/4
    Met1-Asn265 (Met262Leu)
    Accession # Q01860
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    20 µg/mL
    See below
  • Immunoprecipitation
    3 µg/106 cells
    NTera‑2 human testicular embryonic carcinoma cell line, see our available Western blot detection antibodies
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/106 cells
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Chromatin Immunoprecipitation (ChIP)
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. The nanog promoter was detected by standard PCR.
Immunocytochemistry
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 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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
  • Entrez Gene IDs:
    5460 (Human); 18999 (Mouse)
  • Alternate Names:
    3-Oct; Oct-3/4; Oct4; Otf3g; Otf4; Pou5f1
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

17 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Dinaciclib potently suppresses MCL-1 and selectively induces the cell death in human iPS cells without affecting the viability of cardiac tissue
    Authors: K Alsayegh, K Matsuura, H Sekine, T Shimizu
    Sci Rep, 2017;7(0):45577.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  2. Generation of a TLE3 heterozygous knockout human embryonic stem cell line using CRISPR-Cas9
    Stem Cell Res, 2016;17(2):441-443.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC - Fresh
  3. Brain tissue banking for stem cells for our future
    Sci Rep, 2016;6(0):39394.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: IHC Not Specified
  4. Generation of a TLE1 homozygous knockout human embryonic stem cell line using CRISPR-Cas9
    Stem Cell Res, 2016;17(2):430-432.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC Fresh
  5. Semaphorin 3A induces mesenchymal-stem-like properties in human periodontal ligament cells.
    Authors: Wada, Naohisa, Maeda, Hidefumi, Hasegawa, Daigaku, Gronthos, Stan, Bartold, Peter Ma, Menicanin, Danijela, Fujii, Shinsuke, Yoshida, Shinichi, Tomokiyo, Atsushi, Monnouchi, Satoshi, Akamine, Akifumi
    Stem Cells Dev, 2014;23(18):2225-36.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fresh
  6. Effects of antioxidants on the quality and genomic stability of induced pluripotent stem cells.
    Authors: Luo, Lan, Kawakatsu, Miho, Guo, Chao-Wan, Urata, Yoshishi, Huang, Wen-Jing, Ali, Haytham, Doi, Hanako, Kitajima, Yuriko, Tanaka, Takayuki, Goto, Shinji, Ono, Yusuke, Xin, Hong-Bo, Hamano, Kimikazu, Li, Tao-Shen
    Sci Rep, 2014;4(0):3779.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. Pluripotent stem cells derived from mouse and human white mature adipocytes.
    Authors: Jumabay M, Abdmaulen R, Ly A, Cubberly M, Shahmirian L, Heydarkhan-Hagvall S, Dumesic D, Yao Y, Bostrom K
    Stem Cells Transl Med, 2014;3(2):161-71.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  8. Genome-wide chromatin interactions of the Nanog locus in pluripotency, differentiation, and reprogramming.
    Authors: Apostolou E, Ferrari F, Walsh R, Bar-Nur O, Stadtfeld M, Cheloufi S, Stuart H, Polo J, Ohsumi T, Borowsky M, Kharchenko P, Park P, Hochedlinger K
    Cell Stem Cell, 2013;12(6):699-712.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: ChIP
  9. Generation of corneal epithelial cells from induced pluripotent stem cells derived from human dermal fibroblast and corneal limbal epithelium.
    Authors: Hayashi, Ryuhei, Ishikawa, Yuki, Ito, Miyuki, Kageyama, Tomofumi, Takashiba, Kuniko, Fujioka, Tsuyoshi, Tsujikawa, Motokazu, Miyoshi, Hiroyuki, Yamato, Masayuki, Nakamura, Yukio, Nishida, Kohji
    PLoS ONE, 2012;7(9):e45435.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  10. A novel role for an RNA polymerase III subunit POLR3G in regulating pluripotency in human embryonic stem cells.
    Authors: Wong RC, Pollan S, Fong H, Ibrahim A, Smith EL, Ho M, Laslett AL, Donovan PJ
    Stem Cells, 2011;29(10):1517-27.
    Species: Human
    Sample Type: Cell Lysates
    Application: ChIP
  11. Glial cell line-derived neurotrophic factor and endothelial cells promote self-renewal of rabbit germ cells with spermatogonial stem cell properties.
    Authors: Kubota H, Wu X, Goodyear SM, Avarbock MR, Brinster RL
    FASEB J., 2011;25(0):2604-14.
    Species: Rabbit
    Sample Type: Whole Cells
    Application: ICC
  12. SIP1 mediates cell-fate decisions between neuroectoderm and mesendoderm in human pluripotent stem cells.
    Authors: Chng Z, Teo A, Pedersen RA, Vallier L
    Cell Stem Cell, 2010;6(1):59-70.
    Species: Human
    Sample Type: Cell Lysates
    Application: ChIP
  13. Regulation of self-renewal and pluripotency by Sox2 in human embryonic stem cells.
    Authors: Fong H, Hohenstein KA, Donovan PJ
    Stem Cells, 2008;26(8):1931-8.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  14. Differential requirements for hematopoietic commitment between human and rhesus embryonic stem cells.
    Authors: Rajesh</LastName><ForeNam D</Initial, Rajesh D, Chinnasamy N, Mitalipov SM, Wolf DP, Slukvin I, Thomson JA, Shaaban AF
    Stem Cells, 2007;25(2):490-9.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  15. A comparison of NIH-approved human ESC lines.
    Authors: Ware CB, Nelson AM, Blau CA
    Stem Cells, 2006;24(12):2677-84.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  16. Controlled-rate freezing of human ES cells.
    Authors: Ware CB, Nelson AM, Blau CA
    BioTechniques, 2005;38(6):879-80, 882-3.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
Expand to show all 17 Citations
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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 application: Immunocytochemistry/Immunofluorescence.

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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Human/Mouse Oct-3/4 Antibody AF1759
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 ICC/IF Human Anonymous 11/23/2017
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Immunocytochemistry/Immunofluorescence Human/Mouse Oct-3/4 Antibody AF1759
Immunocytochemistry/Immunofluorescence: Human/Mouse Oct-3/4 Antibody [AF1759]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedUmbilical vein endothelial cells
SpeciesHuman

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