Stem Cell Epigenetics
The two hallmark features of stem cells are pluripotency, the ability to differentiate into any mature cell type, and self-renewal, the capacity to undergo indefinite replicative cycles without losing stem cell identity. The general types of stem cells include embryonic stem cells (ESCs), adult stem cells and induced pluripotent stem cells (iPSCs). ESCs are derived from a population of cells in the blastocyst of a pre-implantation embryo called the inner cell mass that can differentiate into any cell type derivative of the three germ layers (endoderm, mesoderm and ectoderm). Adult stem cells can be found throughout the post-embryonic and adult organism, and function primarily in the maintenance, repair and regeneration of tissue and organs. iPSCs are pluripotent stem cells that are artificially derived when adult somatic cells are genetically reprogrammed to an embryonic stem cell phenotype.
Active Motif has a large and expanding collection of tools to help scientists study stem cell biology including antibodies, proteins, and stem cell specific stains.
Live pluripotent stem cell Stain
Active Motif's Stem Cell CDy1 Dye provides a fast and simple method to selectively stain live pluripotent stem cells for identification and sorting using fluorescence microscopy and FACS. Because CDy1 is a live cell dye, it eliminates the need to sacrifice precious stem cell samples for testing, providing a significant advantage over other methods that require cell fixation, such as immunostaining. In comparison to other live cell staining techniques, CDy1 is highly specific for pluripotent stem cells and unlike live alkaline phosphatase staining methods, does not produce background signal in other cells, such as feeder or differentiated cells. Staining is reversible and does not impact cell growth, morphology, or differentiation, making it ideal for stem cell enrichment and sorting, or for performing additional experiments or counterstaining procedures.
Unlike alkaline phosphatase live staining, CDy1 is specific for pluripotent stem cells.
For more detailed information about our Stem Cell CDy1 dye, including instructions for use and ordering information, please click here.
Antibodies for Stem Cell Research
Active Motif offers an extensive collection of antibodies inportant in stem cell research including ChIP-Seq validated antibodies specific for stem cell pluripotency factors. Please see below for additional information about these antibodies.
|KLF5 (Kruppel-like factor 5, IKLF, Basic transcription element-binding protein 2, BTEB2) is a zinc-finger transcription factor that plays a role in cell proliferation, differentiation, cell cycle regulation, angiogenesis, and carcinogenesis. KLF5 is of particular importance in epithelial cells. KLF5 (along with KLF2 and KLF4) is highly expressed in undifferentiated mouse embryonic stem cells and downregulated during early stages of differentiation. Both KLF5 and KLF2 can substitute for KLF4 in the reprogramming of differentiated cells into stem cells. KLF5 also has a proliferative role in cells and the loss of both KLF5 alleles is embryonic lethal.|
|The Nanog (Nanog homeobox) is a transcriptional regulator involved in inner cell mass and embryonic stem (ES) cell proliferation and self-renewal. This protein also imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages, blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes (by similarity), acts as a transcriptional activator or repressor (by similarity), and binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3' (by similarity). When overexpressed, Nanog also promotes cells to enter into S phase and proliferation (by similarity).|
|Oct 4 Antibody|
|Oct 4 (Octamer binding protein-4, POU5F1) is a member of the POU family of transcriptional activators and as such, is critical for early embryogenesis and required for embryonic stem cell pluripotency. Oct-4 is expressed at high levels in undifferentiated cells and ectopic expression of Oct-4 (and several other transcription factors) can induce pluripotency in differentiated cells.|
|Sox 2 antibody|
|The Sox2 (SRY related HMG BOX gene 2) is a DNA binding transcription factor and a member of the SOX family of proteins. Characteristically, SOX proteins have an HMG box that binds DNA. Sox2 forms a complex with Oct-4 and controls the expression of a number of genes involved in embryonic development. Sox2 is also critical for early embryogenesis and embryonic stem cell pluripotency and can thus serve as a stem cell marker. Overexpression of Sox2 (along with Oct-4, KLF4 and c-Myc) can transform mouse fibroblasts into a state resembling embryonic stem cells (ES cells), referred to as Induced Pluripotency. Defects in Sox2 are the cause of microphthalmia syndromic type 3.|
|TCF7L1 / TCF3 antibody|
|TCF3 / TCF7L1 (T-cell factor 3, TCF7L1) is a member of the TCF/LEF family, a component of the Wnt signaling pathway and a dominant downstream effector in embryonic stem cells (ESCs). TCF3 binds to DNA and serves as both a repressor and activator of transcription. TCF3 also brings developmental signals directly to the core regulatory circuitry of ES cells to influence the balance between pluripotency and differentiation. TCF3 transcriptionally represses many genes important for maintaining pluripotency and self-renewal, as well as those involved in lineage commitment and stem cell differentiation. This effect is in part mediated by the corepressor transducin-like enhancer of split 2 (TLE2) and C-terminal Binding Protein (CtBP).|
|The Tet1 (Ten-Eleven Translocation-1; methylcytosine dioxygenase) is a protein that catalyzes the conversion of the modified genomic base 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC), which can lead to cytosine demethylation by either further oxidation via Tet proteins to 5-formylcytosine (5fC) and 5-carboxycytosine (5caC) or deamination into 5-hydroxymethyluracil (5hmU) and then subsequent replacement by unmethylated cytosine by the base excision repair system. Methylation at the C5 position of cytosine bases is an epigenetic modification of the mammalian genome that plays an important role in transcriptional regulation. Tet1 preferentially binds to CpG-rich sequences at promoters of both transcriptionally active and polycomb-repressed genes. By controlling the levels of 5mC and 5hmC at gene promoters, it may regulate the gene expression silencing induced by cytosine methylation and may have a dual function by also repressing the expression of a subset of genes through recruitment of transcriptional repressors to promoters. Tet1 is also involved in the balance between pluripotency and lineage commitment of cells and plays a role in embryonic stem cells maintenance and inner cell mass cell specification.|