Alkaline Phosphatase Live Stain is a stem cell imaging product that allows users to differentially stain pluripotent stem cells (PSCs). The dye is a cell-permeable fluorescent substrate for alkaline phosphatase (AP) that is non-toxic to cells, diffusing out over the course of two hours. The Alkaline Phosphatase Live Stain consists of 50 µL of a 500X fluorescein-based dye in DMSO, sufficient for staining two 24-well plates, four 6 cm dishes or two 10 cm dishes.
This Alkaline Phosphatase Live Stain allows you to:
• Identify pluripotent stem cells with a strong signal above background
• Stain PSCs typically in 20 minutes or less
• Use the first alkaline phosphatase stain marketed that maintains cell viability
Alkaline Phosphatase Pluripotent Stem Cell Detection Method
AP is a phenotypic marker of PSCs, including undifferentiated embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), and embryonic germ cells (EGCs). While AP is expressed in most cell types, its expression is highly elevated in PSCs. AP staining has therefore been used to differentially stain PSCs to easily distinguish them from mouse embryonic fibroblasts (MEFs) used as feeders and parental fibroblasts commonly used in reprogramming experiments.
Screen for Pluripotency Without Sacrificing Precious Colonies
The Alkaline Phosphatase Live Stain is ideal for screening colonies during early stages of the reprogramming workflow since the stain maintains stem cell viability. It can also be used in later stages as a negative selection tool for identifying undifferentiated cells.
Stain Cells Quickly
The Alkaline Phosphatase Live Stain utilizes an easy, non-permanent, cell-viable protocol for identifying PSCs in your experiments. Simply dilute the dye in media, apply to cells, gently wash, and the cells are ready for fluorescent imaging.
For Research Use Only. Not for use in diagnostics procedures.
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![Figure 1: Alkali...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/Web_Figure%201[1]%20635px.jpg","Figure 1: Alkaline Phosphatase (AP) Live Stain shows robust staining in pluripotent stem with minimal background in fibroblasts.","BJ human fibroblast, a common source of somatic cell type used for reprogramming and mitotic inactivated murine embryonic fibroblasts, commonly used as feeders for pluripotent stem cell culture show minimal background with AP Live Stain. Mouse ESC and human ESC grown on MEF feeders show specific staining of the pluripotent stem cell colonies. Human iPSC derived from BJ human fibroblasts using CytoTuneTM Reprogramming Kit shows robust staining with AP Live Stain both on feeders as well feeder-free conditions.");)
![Figure 2: Live A...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/Web_Figure%202[1]%20635px.jpg","Figure 2: Live AP Staining is reversible.","H9 ESC on feeders stained with AP Live Stain show robust staining of the ESC colonies. The green fluorescent signal is lost from the cells by 90 minutes after removal of the dye from the medium.");)
![Figure 3: Viabil...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/Web_Figure%203[1]%20635px.jpg","Figure 3: Viability of cells following staining with AP Live Stain.","Feeder-free H9 ESC stained with Live AP and recovered for 2 h to eliminate the fluorescence stain was assessed for cell health using PrestoBlueTM, a cell vitality dye that results in increase in fluorescence at 590 nm in the presence of metabolically active cells. Cells stained with 4 independent lots of the AP Live Stain showed high signal comparable to control AP Live untreated cells, indicating metabolically active cells. Statistical comparison using ANOVA showed no significant difference between the Live AP treated samples and control cells (P>0.05) but significantly different (P<0.05) compared to cells treated with known cell disruptive agents such as Paraformaldehye (PFA) and Dimethyl sulfoxide (DMSO).");)
![AP Live Stain Wor...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/CO01695-AP-Live-Stain_650pix_d2.jpg","AP Live Stain Workflow","");)
