Recombinant Anti-Oct4 antibody [EPR17980] (ab200834)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17980] to Oct4
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIP-sequencing
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Oct4 antibody [EPR17980]
See all Oct4 primary antibodies -
Description
Rabbit monoclonal [EPR17980] to Oct4 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIP-sequencingmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: NCCIT, Ntera-2/D1 and F9 whole cell lysates. IHC-P: Human dysgerminoma of ovary and spermatocytoma tissues. ICC/IF: NCCIT cells, F9 cells Flow Cyt (intra): F9 cells. IP: F9 whole cell lysate. ChIP-seq: NCCIT cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17980 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Oct4 antibody [EPR17980] (ab208907)
- Alexa Fluor® 647 Anti-Oct4 antibody [EPR17980] (ab209957)
- PE Anti-Oct4 antibody [EPR17980] (ab213198)
- Alexa Fluor® 594 Anti-Oct4 antibody [EPR17980] (ab217250)
- Alexa Fluor® 555 Anti-Oct4 antibody [EPR17980] (ab217743)
- Anti-Oct4 antibody [EPR17980] - BSA and Azide free (ab240358)
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab200834 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 |
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WB |
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 38 kDa).
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IHC-P | (1) |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
Use a concentration of 5 µg/ml.
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IP |
1/30.
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Flow Cyt (Intra) |
1/60.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIP-sequencing |
Use at an assay dependent concentration.
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Notes |
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WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 38 kDa). |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 5 µg/ml. |
IP
1/30. |
Flow Cyt (Intra)
1/60. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIP-sequencing
Use at an assay dependent concentration. |
Target
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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
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 localization
Nucleus. Expressed in a diffuse and slightly punctuate pattern. - Information by UniProt
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Database links
- Entrez Gene: 5460 Human
- Entrez Gene: 18999 Mouse
- Omim: 164177 Human
- SwissProt: Q01860 Human
- SwissProt: P20263 Mouse
- Unigene: 249184 Human
- Unigene: 632482 Human
- Unigene: 646545 Human
see all -
Alternative names
- Octamer binding transcription factor 4 antibody
- MGC22487 antibody
- Oct 3 antibody
see all
Images
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All lanes : Anti-Oct4 antibody [EPR17980] (ab200834) at 1/10000 dilution
Lane 1 : NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate
Lane 2 : NTera-2/D1 (Human malignant pluripotent embryonic carcinoma) whole cell lysate
Lane 3 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 4 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate
Lane 5 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 6 : Mouse testis lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 38 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsLow levels of expression in adult tissues.
Blocking/Dilution Buffer: 5% NFDM/TBST.
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ab200834 staining Oct4 in F9 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab200834 at a 5μg/ml concentration and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1μg/ml concentration, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green) and an anti-mouse AlexaFluor® 594 (ab150120) at 2 μg/ml (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunocytochemistry/Immunofluorescence analysis of F9 (mouse embryonal carcinoma) labelling Oct4 with purified ab200834 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
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All lanes : Anti-Oct4 antibody [EPR17980] (ab200834) at 1/1000 dilution
Lane 1 : Human placenta lysate
Lane 2 : Human fetal brain lysate
Lane 3 : Human testis lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 38 kDa
Exposure time: 3 minutesLow levels of expression in adult tissues
Blocking/Dilution Buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
Immunohistochemical analysis of paraffin-embedded Human dysgerminoma of ovary tissue labeling Oct4 with ab200834 at 1/500 dilution, followed Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human dysgerminoma of ovary tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
Immunohistochemical analysis of paraffin-embedded Human spermatocytoma tissue labeling Oct4 with ab200834 at 1/500 dilution, followed Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human spermatocytoma tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17980] (ab200834)
Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Oct4 with ab200834 at 1/500 dilution, followed Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Human cerebral cortex tissue is a negative control for Oct4. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed F9 (Mouse embyro testicular cancer cell line) cells labeling Oct4 with ab200834 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730;black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
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Oct4 was immunoprecipitated from 1mg of F9 (Mouse embyro testicular cancer cell line) whole cell lysate with ab200834 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200834 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: F9 whole cell lysate 10 µg (Input). Lane 2: ab200834 IP in F9 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200834 in F9 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 5 seconds
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Chromatin was prepared from NCCIT (Human pluripotent embryonic carcinoma cell line) cells. ChIP was performed with 10^7 NCCIT cells and 8 µg of ab200834 [EPR17980]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (25)
ab200834 has been referenced in 25 publications.
- Mei J et al. MACC1 regulates the AKT/STAT3 signaling pathway to induce migration, invasion, cancer stemness, and suppress apoptosis in cervical cancer cells. Bioengineered 13:61-70 (2022). PubMed: 34939526
- Tang Y et al. ING4 Promotes Stemness Enrichment of Human Renal Cell Carcinoma Cells Through Inhibiting DUSP4 Expression to Activate the p38 MAPK/type I IFN-Stimulated Gene Signaling Pathway. Front Pharmacol 13:845097 (2022). PubMed: 35496267
- Tanosaki S et al. Purification of cardiomyocytes and neurons derived from human pluripotent stem cells by inhibition of de novo fatty acid synthesis. STAR Protoc 3:101360 (2022). PubMed: 35516845
- Tiong TY et al. Targeting the SREBP-1/Hsa-Mir-497/SCAP/FASN Oncometabolic Axis Inhibits the Cancer Stem-like and Chemoresistant Phenotype of Non-Small Cell Lung Carcinoma Cells. Int J Mol Sci 23:N/A (2022). PubMed: 35806291
- Xu J et al. UPF1/circRPPH1/ATF3 feedback loop promotes the malignant phenotype and stemness of GSCs. Cell Death Dis 13:645 (2022). PubMed: 35871061