- Datasheet
- Specific References (70)
- Protocols
Overview
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Product nameAnti-Oct4 antibody
See all Oct4 primary antibodies -
DescriptionRabbit polyclonal to Oct4
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Host speciesRabbit
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SpecificityThe antibody reacts with 45 kDa Oct4 (also designated Oct3A) and in a less extent 33 kDa Oct3B from human, mouse, and rat samples.
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Tested applicationsSuitable for: IHC-Fr, WB, IP, ICC/IF, Flow Cyt, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Human, Pig, Macaque monkey
Predicted to work with: Chimpanzee
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Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive).
(Peptide available asab52110) -
Positive control
- Rat kidney tissue lysate can be used as a positive control.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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Storage bufferpH: 7.20
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: 69% PBS, 30% Glycerol, 0.5% BSA -
Concentration information loading... -
PurityProtein A purified
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Purification notesAffinity Purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
Applications
Our Abpromise guarantee covers the use of ab18976 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 |
|---|---|---|
| IHC-Fr | Use a concentration of 20 µg/ml. | |
| WB | Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 45 kDa.Can be blocked with Oct4 peptide (ab52110). | |
| IP | Use a concentration of 20 µg/ml. | |
| ICC/IF | Use at an assay dependent concentration. PubMed: 19736565 | |
| Flow Cyt | Use at an assay dependent concentration. PubMed: 17719011 ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IHC-P | 1/100. Perform enzymatic antigen retrieval before commencing with IHC staining protocol. |
Target
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FunctionTranscription 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.
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Tissue specificityExpressed 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.
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Sequence similaritiesBelongs to the POU transcription factor family. Class-5 subfamily.
Contains 1 homeobox DNA-binding domain.
Contains 1 POU-specific domain. -
Developmental stageHighly expressed in undifferentiated embryonic stem cells and expression decreases gradually after embryoid body (EB) formation.
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DomainThe POU-specific domain mediates interaction with PKM2.
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Post-translational
modificationsSumoylation 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 localizationNucleus. Expressed in a diffuse and slightly punctuate pattern.
- Information by UniProt
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Database links
- Entrez Gene: 744263 Chimpanzee
- Entrez Gene: 5460 Human
- Entrez Gene: 18999 Mouse
- Entrez Gene: 100127461 Pig
- Entrez Gene: 294562 Rat
- Omim: 164177 Human
- SwissProt: Q7YR49 Chimpanzee
- SwissProt: Q01860 Human
see all -
Alternative names
- Octamer binding transcription factor 4 antibody
- MGC22487 antibody
- Oct 3 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody (ab18976)Image from Hassiotou et al., Front Oncol. 2013 3:79. Figure 1A doi: 10.3389/fonc.2013.00079. eCollection 2013.Immunohistochemistry analysis of paraffin-embedded human breast tissues (5 µm thick) labeling Oct4 with ab18976 at 1/100 dilution.
Tissues were rehydrated in deionized water and incubated in PBS prior to per meabilization in 0.1% PBS-TritonX 100 and overnight incubation with ab18976 in a humid chamber.Washing in PBS and secondary antibody (Alexa Fluor®) incubation for 2h was followed by a final wash and mounting.
Blue: DAPI stain for nuclei.
Minimal and mostly cytoplasmic expression was seen in the normal resting breast (left panel). During normal lactation (right panel), OCT4 was upregulated and expressed in both the nucleus and cytoplasm.
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Western blot - Anti-Oct4 antibody (ab18976)This image is courtesy of an anonymous AbreviewAnti-Oct4 antibody (ab18976) at 1/500 dilution + mouse embryonic stem cell whole cell lysate at 100 µg/ml
Secondary
HRP conjugated goat anti-rabbit
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 45 kDa
Additional bands at: 25 kDa (possible non-specific binding), 60 kDa (possible non-specific binding)
Exposure time: 1 minuteThe blot was blocked with 5% milk for 1 hour at RT and inbubated with the primary antibody for 16 hours at 4°C.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody (ab18976)Image from Hassiotou et al., Front Oncol. 2013 3:79. Figure 1A doi: 10.3389/fonc.2013.00079. eCollection 2013.Immunohistochemical analysis of paraffin embedded Human breast tissues (5 µm thick) labeling Oct4 with ab18976 at 1/100 to 1/200 dilution. Heat induced epitope retrieval was performed at 100°C for 30 min. Slides were counterstained with hematoxylin.
Minimal and mostly cytoplasmic expression was seen in the normal resting breast (left panel). During normal lactation (right panel), OCT4 was upregulated and expressed in both the nucleus and cytoplasm.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody (ab18976)This image is courtesy of an anonymous Abreviewab18976 at 1/100 dilution staining Oct-4 in mouse brain tissue by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were paraformaldehyde fixed prior to blocking in 5% BSA for 1 hour at 25°C and then incubated with ab18976 for 12 hours at 4°C. A Cy3 conjugated donkey polyclonal to rabbit Ig, diluted 1/500, was used as the secondary antibody.
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Western blot - Anti-Oct4 antibody (ab18976)Anti-Oct4 antibody (ab18976) at 0.5 µg/ml + Lysates prepared from rat kidney tissue.
Predicted band size: 45 kDaThe antibody reacts with 45 kDa Oct4 (also designated Oct-3A) and in a less extent 33 kDa Oct-3B from human, mouse, and rat samples.
Protocols
References
This product has been referenced in:
- Su X et al. Embryonic lethality in mice lacking Trim59 due to impaired gastrulation development. Cell Death Dis 9:302 (2018). Read more (PubMed: 29467473) »
- Balahmar RM et al. Identification and characterisation of NANOG+/ OCT-4high/SOX2+ doxorubicin-resistant stem-like cells from transformed trophoblastic cell lines. Oncotarget 9:7054-7065 (2018). Read more (PubMed: 29467949) »
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