Product FAQ

How do I visualize cells stained with Alkaline Phosphatase Live Stain?

Answer

After 20-30 minutes, remove the AP Live Stain and wash the cells twice for 5 minutes each with DMEM/F-12 (Cat. No. 10565-018). Following the final wash, add fresh DMEM/F-12 and visualize the stained colonies under fluorescent microscopy using a standard FITC filter. Images should be captured within 10 to 30 minutes following the removal of the dye and the most robust fluorescent colonies should be marked for selection and expansion. Since the fluorescent signal leeches out of the cells and into the surrounding medium as the stain is turned over, we strongly encourage that you visualize and capture images immediately following the final wash for optimal signal detection.

Answer Id: E12567

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Product FAQ

How do I prepare the StemPRO hESC SFM complete medium?

Answer

We recommend to thaw the supplement in the 37°C water bath, aliquot and freeze at -5°C to -20°C. Thaw each aliquot only one additional time. To make 100 ml complete medium, mix 90.8 ml DMEM/F-12 with GlutaMax matrix, 2 ml StemPRO hESC supplement, 7.2 ml 25% BSA solution, 80 ul bFGF (at 10 ug/ml), and 182 ul 2-mercaptoethanol (at 55mM). Store at 2°C-8°C until ready to use, but use the same day.

Answer Id: E5304

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Product FAQ

How can I prevent poor or dim staining when using Alkaline Phosphatase Live Stain?

Answer

Serum or serum replacement components in the growth medium may cause background and this can result in poor or dim staining. After the removal of the growth medium, gently wash the culture with pre-warmed DMEM/F-12 (Cat. No. 10565-018) for 5 minutes. Aspirate and repeat once before adding the AP Live Stain. To further decrease background staining, perform 3 separate washes of 5 minutes each with DMEM/F-12 for a total of 15 minutes following the staining, and visualize immediately.

Answer Id: E12574

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Citations & References

Blocking HES1 expression initiates GABAergic differentiation and induces the expression of p21(CIP1/WAF1) in human neural stem cells.

  • Authors: Kabos Peter; Kabosova Andrea; Neuman Toomas;
  • Journal: J Biol Chem (2002) 277:8763-6
  • PubMed ID: 11809764

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