Oct-4 antibody (pAb)

Aliases: POU5F1, OCT3, OTF3, OTF4, OTF-3, Oct-3
Catalog No: 39811 Format: 100 µl $445 Buy
Catalog No: 39812 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Affinity Purified
Host:
Rabbit
Molecular Weight:
45 kDa
Reactivity:
Human, Mouse

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes


Validated Applications:
ChIP: 10 µl per ChIP
ChIP-Seq: 10 µl each
ICC/IF: 1:200 dilution
WB: 1:1,000 - 1:5,000 dilution

ChIP-Seq validation was performed by Active Motif's Epigenetics Services and the complete data set is available in the UCSC Genome Browser by clicking here.

Published Applications:
ChIP
WB
IF

Immunogen

This Oct-4 antibody was raised against a peptide derived from human Oct-4.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 
Oct-4 antibody (pAb) tested by ChIP-Seq.

Oct-4 antibody (pAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from undifferentiated hESC cells and 7 µl of antibody. ChIP DNA was sequenced on the Illumina HiSeq and11 million sequence tags were mapped to identify Oct-4 binding sites. The image shows binding across a region of chromosome 11. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Oct-4 antibody (pAb) tested by immunofluorescence.

Oct-4 antibody (pAb) tested by Immunofluorescence
Mouse embryonic stem cells (mESCs) grown on mouse embryonic fibroblast feeder cells (MEFs) were fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were then permeabilized and blocked by incubating with Blocking Solution containing 5% serum/0.1% Triton X-100 in D-PBS for 2 hours at room temperature. Cells were then incubated with Oct-4 antibody (Catalog No. 39811, red) at 1:200 dilution overnight at 4°C, washed with D-PBS, and incubated for 2 hours at room temperature with goat anti-mouse Cy3 secondary antibody at 1:250 dilution. Nuclei were stained with DAPI (blue). Cells were visualized using a Zeiss fluorescent microscope at 20X magnification. Images show that Oct-4 antibody specifically stains mESC colonies and does not stain MEFs. Absence of Oct-4 staining in a subset of cells within the colonies suggests differentiation. Scale bars, 100 μm.

Oct-4 antibody (pAb) tested by Western blot.

Oct-4 antibody tested by Western blot.
Detection of Oct-4 by Western blot. P19 cell nuclear extract (25 µg) probed with Oct-4 antibody (pab) (1:1,000 dilution).

Background

Oct-4 (Octamer binding protein-4, POU5F1) is a member of the POU family of transcriptional activators. Oct-4 is critical for early embryogenesis and required for embryonic stem cell pluripotency. Oct-4 is expressed at high levels in undifferentiated cells and ectopic expression of Oct-4 (and several other transcription factors) can induce pluripotency in differentiated cells.

What are the Epigenetic Determinants of Stem Cell Identity?

Positive Control

Active Motif's P19 nuclear extract (Catalog No. 36074) can be used as a positive control for Oct-4 antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Oct-4 antibody (pAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Oct-4 antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting