Overview

  • Product name
  • Description
    Goat polyclonal to Oct4
  • Host species
    Goat
  • Specificity
    According to the UniProt database Oct4 is mainly localized in the nucleus but can also occur in the cytoplasm: UniProtKB - Q01860
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Oct4 aa 300 to the C-terminus (C terminal).
    (Peptide available as ab20650)

  • Positive control
    • Human or Mouse Embryonic Stem Cells

Properties

Applications

Our Abpromise guarantee covers the use of ab27985 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
Flow Cyt Use at an assay dependent concentration.

ab37373 - Goat polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 1 - 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 39 kDa).

Target

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

Images

  • ab27985 recognizes Oct4 in human embryonic stem cells. The upper panel shows DAPI staining while the lower panel shows ab27985 staining.

    See Abreview

  • All lanes : Anti-Oct4 antibody (ab27985) at 1 µg/ml

    Lane 1 : Human Embryonic Stem Cell Lysate
    Lane 2 : Mouse Embryonic Stem Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat IgG secondary antibody (ab28445) at 1/10000 dilution

    Predicted band size: 39 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?



    The antibody recognised a band corresponding to Oct4 in mouse and human embryonic stem cell lysates.

  • ab27985 staining Oct4 in human embryonic carcinoma cells by Flow Cytometery. Cells were fixed in paraformaldehyde and permeabilized in saponin buffer. The primary antibody was diluted 1:500 (PBS + 10% BSA) and incubated with sample for 1 hour at 4°C. A FITC conjugated Donkey polyclonal to  goat IgG, diluted 1:200 was used as secondary. The red histogram is secondary only control whilst green and blue repersents Oct4 expression in untreated and partially differentiated with 10nM Trans-Retinoic acid for 14 days. 

    See Abreview

  • ab27985 recognizes Oct4 in mouse embryonic stem cells. DAPI staining is shown in the top panel while ab27985 staining is shown in the lower panel.

    See Abreview

References

This product has been referenced in:
  • Peng Y  et al. Human embryonic stem cells extracellular vesicles and their effects on immortalized human retinal Müller cells. PLoS One 13:e0194004 (2018). Read more (PubMed: 29538408) »
  • Orsztynowicz M  et al. Changes in chromosome territory position within the nucleus reflect alternations in gene expression related to embryonic lineage specification. PLoS One 12:e0182398 (2017). Read more (PubMed: 28767705) »
See all 44 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Mouse Cell (Embryonic Stem Cell)
Permeabilization
Yes - 0.2% Triton X
Specification
Embryonic Stem Cell
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 01 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
Sample
Mouse Tissue sections (mice embryo 14.5 dpc)
Specification
mice embryo 14.5 dpc
Permeabilization
Yes - Tween-20
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 10 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
Sample
Dog Tissue sections (MDCK pellets in paraffin)
Specification
MDCK pellets in paraffin
Permeabilization
Yes - Tween-20
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 04 2015

Answer

Thank you for the details provided. I am sorry you are having difficulty with these products.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab27985 and ab96916.

1) Increase the Triton X to 0.25% for 10 min to ensure permeabilization so the nuclear stain can be seen fully

2) Add a no primary control to see if the background is from the secondary. If it is, you may wish to use 10% rabbit serum instead of 5% BSA to block background from the secondary.

3) Try increasing primary antibody incubation to ON at 4C to ensure specific signal is strong.

4) Add extra washes after primary and secondary to reduce background.


Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (embryonic stem cell)
Loading amount
1e+006 cells
Specification
embryonic stem cell
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Aug 14 2012

Answer

Thank you for contacting Abcam.

Based on the information we have for this antibody, it may be that it is not suitable for IHC-Fr on sections. However based on the information you have provided, you could try to increase the primary antibody concentration to a 1/50 dilution and incubate overnight at 4C.

I have also attached the Abcam protocols book that contains more useful information on troubleshooting with IHC-Fr.

We do have a few other Oct4 antibodies that we guarantee to work in IHC-Fr on mouse sections and I have placed links to these below:

https://www.abcam.com/Oct4-antibody-ChIP-Grade-ab19857.html
https://www.abcam.com/Oct4-antibody-ab18976.html



If there is anything else I can help you with, please let me know.

Read More

Question
Answer

Thank you for your call today and for letting us know about the trouble with these antibodies.

Please keep me updated about the results with these antibodies after trying the alterations that we discussed. Here is the article that I mentioned about performing heat retrieval on frozen tissue sections-

http://jhc.sagepub.com/content/53/11/1421.full

This article isn't specific for Brdu pre-treatment but it might be a helpful resource for you if you'd prefer to try a heat denaturation before using the HCl method. I've found another article (attached) which will useful for heat denaturation as well.

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More

Answer

Thank you for contacting us. The closest match we have to ab28445 is ab96930. This is an anti-goat IgG, raised in rabbit, conjugated to DyLight® 650, which has an excitation maximum at 654nm and an emission maximum at 673nm. The Alexa Fluor 680 fluorophore of ab28445, on the ohter hand, has an excitation maximum at 680nm and emission at 702nm. Both are in the far red range of the visible light spectrum. Click here (or use the following: https://www.abcam.com/index.html?datasheet=96930). Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (E12.5 testis)
Specification
E12.5 testis
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton or 1% Triton
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 25 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis)
Specification
Testis
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 08 2011

1-10 of 15 Abreviews or Q&A

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