Overview

  • Product name

    Human Oct4 ELISA Kit
    See all Oct4 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Cell extract 8 9.8%
  • Sample type

    Cell culture extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    596 pg/ml
  • Range

    1.76 ng/ml - 112.5 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 102 % - %

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Oct4 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Oct4 protein in human cell extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Oct4 is a transcription factor that plays a key role in the self-renewal of undifferentiated embryonic stem cells. Up or down regulation of this protein leads to aberrant differentiation. Induction of Oct4, in combination with other transcription factors, is used to reprogram cells and induce a pluripotent stem cell-like state. Oct4 is associated with the plasticity of cancer stem cells.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BR 1 x 6ml
    Human Oct4 Lyophilized Recombinant Protein 2 vials
    10X Human Oct4 Capture Antibody 1 x 600µl
    10X Human Oct4 Detector Antibody 1 x 600µl
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Transcription factor that binds to the octamer motif (5'-ATTTGCAT-3'). Forms a trimeric complex with SOX2 on DNA and controls the expression of a number of genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206. Critical for early embryogenesis and for embryonic stem cell pluripotency.
  • Tissue specificity

    Expressed in developing brain. Highest levels found in specific cell layers of the cortex, the olfactory bulb, the hippocampus and the cerebellum. Low levels of expression in adult tissues.
  • Sequence similarities

    Belongs to the POU transcription factor family. Class-5 subfamily.
    Contains 1 homeobox DNA-binding domain.
    Contains 1 POU-specific domain.
  • Developmental stage

    Highly expressed in undifferentiated embryonic stem cells and expression decreases gradually after embryoid body (EB) formation.
  • Domain

    The POU-specific domain mediates interaction with PKM2.
  • Post-translational
    modifications

    Sumoylation enhances the protein stability, DNA binding and transactivation activity. Sumoylation is required for enhanced YES1 expression.
    Ubiquitinated; undergoes 'Lys-63'-linked polyubiquitination by WWP2 leading to proteasomal degradation.
  • Cellular localization

    Nucleus. Expressed in a diffuse and slightly punctuate pattern.
  • Information by UniProt
  • Alternative names

    • Octamer binding transcription factor 4
    • MGC22487
    • Oct 3
    • Oct 4
    • Oct-3
    • Oct-4
    • OCT3
    • Oct4
    • Octamer binding protein 3
    • Octamer binding protein 4
    • Octamer binding transcription factor 3
    • Octamer-binding protein 3
    • Octamer-binding protein 4
    • Octamer-binding transcription factor 3
    • OTF 3
    • OTF 4
    • OTF-3
    • OTF3
    • OTF4
    • PO5F1_HUMAN
    • POU class 5 homeobox 1
    • POU domain class 5 transcription factor 1
    • POU domain transcription factor OCT4
    • POU domain, class 5, transcription factor 1
    • POU-type homeodomain-containing DNA-binding protein
    • POU5F1
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab235653 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Oct4 were measured in duplicate and interpolated from the Oct4 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Oct4 concentration was determined to be 26.7 ng/mL in NCCIT cell extract.

Protocols

References

ab235653 has not yet been referenced specifically in any publications.

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