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Vector red alkaline phosphate substrate kit

Manufactured by Vector Laboratories
Sourced in United States

The Vector Red Alkaline Phosphate Substrate Kit is a laboratory reagent used for the detection and visualization of alkaline phosphatase enzyme activity. It provides a chromogenic substrate that undergoes a color change reaction in the presence of the alkaline phosphatase enzyme, allowing for the identification and localization of enzyme-labeled targets.

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2 protocols using vector red alkaline phosphate substrate kit

1

Immunocytochemical Characterization of Pluripotent Stem Cells

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Alkaline phosphatase staining was performed with the Vector Red Alkaline Phosphate Substrate Kit (Vector Laboratories, Burlingame, CA). For immunostaining, cells were fixed in 4% paraformaldehyde with 1% sucrose in phosphate buffered saline (PBS) (Invitrogen) for 15 min at room temperature. The cell membranes were then permeabilized with 0.5% TX-100 in PBS-T (Sigma-Aldrich), blocked in donkey serum (Sigma-Aldrich) and incubated with donkey serum containing primary antibodies including rabbit anti-OCT4 (Merck Millipore), rabbit anti-SOX2 (Abcam, San Francisco, CA) and rabbit anti-NANOG (Merck Millipore) at 1:100 dilutions, washed in PBS-T, and then incubated with Alexa Fluor 594 conjugated goat anti-rabbit secondary antibody (Cell Signaling Technology, Danvers, MA) at a 1:500 dilution. The cell nuclei were counterstained with DAPI. Fluorescent images were taken using a Zeiss or Nikon fluorescence microscope.
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2

Alkaline Phosphate Staining of PC-Enriched Cells

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The cells from PC-enriched supernatants were plated in a 12-well plate (3 × 10 5 cells/well) in MEM containing 10% FBS and P/S and stained using a Vector Red Alkaline phosphate substrate kit (Vector Laboratories, Burlingame, CA, USA) according to the manufacturer's protocol.
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