Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Immunocytochemistry [1]
- Other assay [2]
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- Product number
- 41-5841-80 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- OCT3/4 Monoclonal Antibody (EM92), eFluor™ 570, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The EM92 monoclonal antibody reacts with mouse and human Oct3/4, encoded by the Pou5F1 gene. Oct3/4 is a POU domain-containing transcription factor that is critical for maintaining embryonic stem (ES) and induced pluripotent stem (iPS) cells in a pluripotent state, and is expressed by ES, embryonic germ cells and embryonic carcinoma cell lines. In cells of the inner cell mass (ICM), reduction of Oct3/4 expression causes dedifferentiation to trophoectoderm, whereas increased expression results in differentiation to mesoderm and primitive endoderm. Oct3/4 regulates the expression of several genes, including FGF-4, UTF1, Sox2, Fbx15, Rex1 and osteopontin through distinct mechanisms. Furthermore, Oct3/4 frequently acts synergistically with Sox2 to regulate target gene expression, as is the case with FGF-4. It has been demonstrated that Oct3/4 expression in ES cells can be negatively regulated by either treatment with retinoic acid, or by removal of leukemia-inhibitory factor (LIF).
- Antibody clone number
- EM92
- Concentration
- 0.2 mg/mL
Submitted references Comparative analysis of CI- and CIV-containing respiratory supercomplexes at single-cell resolution.
Pressure-Driven Mitochondrial Transfer Pipeline Generates Mammalian Cells of Desired Genetic Combinations and Fates.
Bertan F, Wischhof L, Scifo E, Guranda M, Jackson J, Marsal-Cots A, Piazzesi A, Stork M, Peitz M, Prehn JHM, Ehninger D, Nicotera P, Bano D
Cell reports methods 2021 May 24;1(1):100002
Cell reports methods 2021 May 24;1(1):100002
Pressure-Driven Mitochondrial Transfer Pipeline Generates Mammalian Cells of Desired Genetic Combinations and Fates.
Patananan AN, Sercel AJ, Wu TH, Ahsan FM, Torres A Jr, Kennedy SAL, Vandiver A, Collier AJ, Mehrabi A, Van Lew J, Zakin L, Rodriguez N, Sixto M, Tadros W, Lazar A, Sieling PA, Nguyen TL, Dawson ER, Braas D, Golovato J, Cisneros L, Vaske C, Plath K, Rabizadeh S, Niazi KR, Chiou PY, Teitell MA
Cell reports 2020 Dec 29;33(13):108562
Cell reports 2020 Dec 29;33(13):108562
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry on fixed and permeabilized F9 cells using 5 µg/mL Rat IgG2a K Isotype Control eFluor® 570 (left) or 5 µg/mL Anti-Human/Mouse OCT3/4 eFluor® 570 (right). Nuclei are stained with DAPI. Colocalization of OCT3/4 and DAPI appears pink.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3. SIMR Fibroblasts Can Be Reprogrammed (A) Native BJ and SIMR fibroblasts reprogrammed to iPSCs with TRA-1-60 + clones counted by microscopy. Data are the means of biological duplicates. Data for BJ fibroblast control are the same data as in Figure S3A . (B) Flow cytometry of pluripotency biomarkers SOX2 and OCT3/4, and fibroblast biomarker CD44. Immunostained samples are shown in color with isotype negative controls in gray. Representative data for native BJ fibroblasts and BJ-iPSCs, and for BJ rho0+PBMC1-iPSC cells. Data for the native BJ fibroblasts and BJ-iPSCs shown here are the same as in Figure S3B . (C) Representative phase contrast and IF microscopy images of native BJ fibroblast (negative control), BJ-iPSC (positive control), and three BJ rho0+PBMC1-iPSC clones immunostained for pluripotency biomarkers SSEA-4 and OCT4. Scale bars, 100 mum. (D) OCR measurements for ~1.5 x 10 4 native BJ-iPSCs and BJ rho0+PBMC1-iPSC clones 1, 2, and 11. Data for BJ-iPSC control are the same as in Figure S3D . Data are the means +- SD of four technical replicates. Statistical significance by unpaired, two-tailed Student''s t test. *p 0.05; **p 0.01 See also Figure S1 and Table S2 .
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4 CI*CIV-SC remodeling occurs during iPSC differentiation (A) OCR measurement of iPSCs and iPSC-derived smNPCs using a conventional Seahorse protocol (n = 3). (B) Mitochondrial spare respiratory capacity in human iPSCs and smNPCs. Percentage is relative to their respective basal OCRs (Student's t test, ****p < 0.0001). (C) Basal OCR in iPSCs and smNPCs (Student's t test, ****p < 0.0001). (D) Immunoblots using antibodies against OXPHOS system components in iPSCs and smNPCs. Densitometry is relative to iPSCs and reported as mean +- SEM (n = 3, Student's t test, ***p < 0.001, *p < 0.05). (E) BN-PAGE and corresponding immunoblots using antibodies against MTCO1 and NDUFB8. (F) Representative pictures of iPSCs and smNPCs stained with DAPI (nucleus, blue), TOM20 (mitochondria, green), and PLA (CI*CIV-SCs, red). Oct3 and nestin staining (both in gray) was used as markers of pluripotency and differentiation, respectively. Scale bar, 5 mum. (G) Quantification of PLA dots normalized to mitochondrial area in iPSCs and smNPCs (n = 3; iPSCs = 20 cells; smNPCs = 17 cells; Student's t test, ***p < 0.001). (H) OCR measurement of WT (P) and two independent clones of NDUFS4 KO iPSCs (KO 1 and KO 2 ). (I) Basal OCR and mitochondrial spare capacity of WT and NDUFS4 KO iPSCs (fold change, relative to WT; two-way RM ANOVA, ***p < 0.001, *p < 0.05; ns, not significant). (J) Immunoblot analysis of homogenates from parental and NDUFS4 KO iPSCs using antibodies against ETC subunits (CI, NDUFS4,