Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20

Mouse monoclonal IgG2b antibody against human OCT4 (OCT3)
概要
The 3A2A20 antibody reacts with human OCT4 (octamer-binding transcription factor 4; also known as OCT3 and OCT3/4), an ~40 kDa homeodomain transcription factor belonging to the POU family, which is expressed in undifferentiated human embryonic stem (ES), induced pluripotent stem (iPS), embryonal carcinoma (EC) and embryonic germ (EG) cells. OCT4 binds to the octamer motif (5'-ATTTGCAT-3') and plays a key role in maintaining cells in a pluripotent state by interacting with other transcription factors such as SOX2 to regulate the expression of several genes, including FBX15, FGF-4, REX1, SOX2 and osteopontin. Levels of OCT4 are down-regulated during differentiation and it has thus emerged as a useful marker of pluripotency in stem cells, as well as a marker for certain human malignant germ cell tumours. Expression of OCT4 together with other transcription factors has been used to reprogram somatic cells into iPS cells. Multiple isoforms of OCT4 have been observed and in humans at least two are functionally active.
Subtype
Primary Antibodies
Target Antigen
OCT4 (OCT3)
Alternative Names
OCT-3, OCT3, OCT-4, octamer-binding transcription factor 4, POU domain class 5 transcription factor 1, POU5F1
Reactive Species
Human
Conjugation
Alexa Fluor 488, PE, Unconjugated
Host Species
Mouse
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Flow Cytometry, Immunocytochemistry, Immunofluorescence, Western Blotting
Area of Interest
Stem Cell Biology
Clone
3A2A20
Gene ID
5460
Isotype
IgG2b, kappa
数据及文献

Data

Data for Alexa Fluor® 488-Conjugated

Figure 1. Data for Alexa Fluor® 488-Conjugated

(A) Flow cytometry analysis of ES cells cultured with mTeSR™1 on Corning® Matrigel®. The ES cells (filled histogram) or HT1080 fibrosarcoma cells (negative control; dashed line histogram) were fixed and labeled with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, Alexa Fluor® 488. Labeling of the ES cells with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, Alexa Fluor® 488 (Catalog #60072AD) is shown (solid line histogram). (B) Human ES cells were cultured with TeSR™-E8™ on glass coverslips coated with Vitronectin XF™ (Catalog #07180), then fixed and labeled with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, Alexa Fluor® 488. Inset shows labeling of human ES cells with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, Alexa Fluor® 488. (C) DAPI counterstaining of the cells shown in figure (B); nuclear localization of the OCT4 (OCT3) marker is evident.

Data for PE-Conjugated

Figure 2. Data for PE-Conjugated

(A) Flow cytometry analysis of human ES cells cultured with mTeSR™1 on Corning® Matrigel®. The ES cells (filled histogram) or HT1080 fibrosarcoma cells (negative control; dashed line histogram) were fixed and labeled with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, PE. Labeling of the ES cells with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, PE (Catalog #60072PE) is shown (solid line histogram). (B) Human ES cells were cultured with TeSR™-E8™ on glass coverslips coated with Vitronectin XF™ (Catalog #07180), then fixed and labeled with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, PE. Inset shows labeling of human ES cells with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, PE. (C) DAPI counterstaining of the cells shown in figure (B); nuclear localization of the OCT4 (OCT3) marker is evident.

Data for Unconjugated

Figure 3. Data for Unconjugated

(A) Human induced pluripotent stem (iPS) cells were cultured with TeSR™-E8™ on glass coverslips coated with Vitronectin XF™ (Catalog #07180), then fixed and stained with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, followed by goat anti-mouse IgG, FITC. Inset shows cells labeled with a mouse IgG2b, kappa isotype control antibody followed by goat anti-mouse IgG, FITC.
(B) Flow cytometry analysis of human embryonic stem (ES) cells cultured with mTeSR™1 on Corning® Matrigel®. The ES cells (filled histogram) or HT1080 fibrosarcoma cells (negative control, dashed line histogram) were fixed and labeled with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, followed by goat anti-mouse IgG, FITC. Labeling of the ES cells with a mouse IgG2b, kappa isotype control antibody followed by goat anti-mouse IgG, FITC is shown (solid line histogram). (C) Flow cytometry analysis of human iPS cells cultured with TeSR™-E8™ on Vitronectin XF™. The cells were fixed and labeled with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20, followed by goat anti-mouse IgG, FITC (filled histogram) or a mouse IgG2b, kappa isotype control antibody followed by goat anti-mouse IgG, FITC (solid line histogram). (D) Western blot analysis of denatured/reduced cell lysates with Anti-Human OCT4 (OCT3) Antibody, Clone 3A2A20. Lane 1, human ES cells cultured with mTeSR™1 on Corning® Matrigel®, lane 2 (negative control), mouse E13.5 neural progenitor cells cultured with NeuroCult™ Proliferation Kit (Mouse, Catalog #05702).

Publications (2)

Scientific reports 2014 NOV Trend of telomerase activity change during human iPSC self-renewal and differentiation revealed by a quartz crystal microbalance based assay. Zhou Y et al.
Abstract
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Abstract

Telomerase plays an important role in governing the life span of cells for its capacity to extend telomeres. As high activity of telomerase has been found in stem cells and cancer cells specifically, various methods have been developed for the evaluation of telomerase activity. To overcome the time-consuming procedures and complicated manipulations of existing methods, we developed a novel method named Telomeric Repeat Elongation Assay based on Quartz crystal microbalance (TREAQ) to monitor telomerase activity during the self-renewal and differentiation of human induced pluripotent stem cells (hiPSCs). TREAQ results indicated hiPSCs possess invariable telomerase activity for 11 passages on Matrigel and a steady decline of telomerase activity when differentiated for different periods, which is confirmed with existing golden standard method. The pluripotency of hiPSCs during differentiation could be estimated through monitoring telomerase activity and compared with the expression levels of markers of pluripotency gene via quantitative real time PCR. Regular assessment for factors associated with pluripotency or stemness was expensive and requires excessive sample consuming, thus TREAQ could be a promising alternative technology for routine monitoring of telomerase activity and estimate the pluripotency of stem cells.
Glycobiology 2014 MAY Evidences for the involvement of cell surface glycans in stem cell pluripotency and differentiation Alisson-Silva F et al.
Abstract
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Abstract

Induced pluripotent stem (iPS) cells are somatic cells that have been reprogrammed to a pluripotent state via the introduction of defined transcription factors. Although iPS is a potentially valuable resource for regenerative medicine and drug development, several issues regarding their pluripotency, differentiation propensity and potential for tumorigenesis remain to be elucidated. Analysis of cell surface glycans has arisen as an interesting tool for the characterization of iPS. An appropriate characterization of glycan surface molecules of human embryonic stem (hES) cells and iPS cells might generate crucial data to highlight their role in the acquisition and maintenance of pluripotency. In this study, we characterized the surface glycans of iPS generated from menstrual blood-derived mesenchymal cells (iPS-MBMC). We demonstrated that, upon spontaneous differentiation, iPS-MBMC present high amounts of terminal $\$-galactopyranoside residues, pointing to an important role of terminal-linked sialic acids in pluripotency maintenance. The removal of sialic acids by neuraminidase induces iPS-MBMC and hES cells differentiation, prompting an ectoderm commitment. Exposed $\$-galactopyranose residues might be recognized by carbohydrate-binding molecules found on the cell surface, which could modulate intercellular or intracellular interactions. Together, our results point for the first time to the involvement of the presence of terminal sialic acid in the maintenance of embryonic stem cell pluripotency and, therefore, the modulation of sialic acid biosynthesis emerges as a mechanism that may govern stem cell differentiation.
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