Anti-Oct4 抗体 [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, ChIC/CUT&RUN-seq
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Oct4 antibody [EPR17980]
Oct4 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR17980] to Oct4 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIP-sequencing, ChIC/CUT&RUN-seqmore details -
種交差性
交差種: Mouse, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- 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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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR17980 -
アイソタイプ
IgG -
研究分野
関連製品
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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
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab200834の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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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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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5 µg |
特記事項 |
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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. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5 µg |
ターゲット情報
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機能
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. -
組織特異性
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. -
配列類似性
Belongs to the POU transcription factor family. Class-5 subfamily.
Contains 1 homeobox DNA-binding domain.
Contains 1 POU-specific domain. -
発生段階
Highly expressed in undifferentiated embryonic stem cells and expression decreases gradually after embryoid body (EB) formation. -
ドメイン
The POU-specific domain mediates interaction with PKM2. -
翻訳後修飾
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. -
細胞内局在
Nucleus. Expressed in a diffuse and slightly punctuate pattern. - Information by UniProt
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参照データベース
- 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 -
別名
- Octamer binding transcription factor 4 antibody
- MGC22487 antibody
- Oct 3 antibody
see all
画像
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 NCCIT (Human pluripotent embryonic carcinoma cell line) cells and 5 µg of ab200834 [EPR17980]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The ChIP data was conducted on chromatin prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8 µg of ab200834. 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.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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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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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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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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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (25)
ab200834 は 25 報の論文で使用されています。
- 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