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Alkaline phosphatase live stain kit

Manufactured by Thermo Fisher Scientific
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The Alkaline Phosphatase Live Stain kit is a reagent used to visually detect and identify live cells expressing alkaline phosphatase, an enzyme involved in various cellular processes. The kit provides a fluorescent substrate that is cleaved by the enzyme, allowing the visualization of alkaline phosphatase-positive cells.

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Lab products found in correlation

Stainalive ssea 4 mouse anti human antibody, by ReproCELL (2 mentions) Bz x700 series microscope, by Keyence (2 mentions) Cytotune ips 2.0 sendai reprogramming kit, by Thermo Fisher Scientific (1 mentions) Nestin, by Merck Group (1 mentions) Foxa2, by Santa Cruz Biotechnology (1 mentions) Nanog, by Santa Cruz Biotechnology (1 mentions) Scid beige mice, by Charles River Laboratories (1 mentions) Hnf3β, by Santa Cruz Biotechnology (1 mentions) Prolong anti fade reagents, by Thermo Fisher Scientific (1 mentions) Hesc lines h1 and h9, by WiCell (1 mentions) Click it edu alexa fluor 594 imaging kit, by Thermo Fisher Scientific (1 mentions) Sb431542, by Selleck Chemicals (1 mentions) Accutase, by Thermo Fisher Scientific (1 mentions) Ldn193189, by Selleck Chemicals (1 mentions) Y 27632, by Selleck Chemicals (1 mentions) Matrigel, by BD (1 mentions)

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Alkaline phosphatase (63 citations) Alkaline Phosphatase Live Stain (2 citations)

5 protocols using alkaline phosphatase live stain kit

1

Alkaline Phosphatase Staining of iPSCs

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Alkaline phosphatase staining was performed using an Alkaline Phosphatase Live Stain Kit (Thermo Fisher, Waltham, MA, USA) and following the manufacturer’s instructions. IPSCs colonies were visualized with Olympus iX73 (Olympus, Tokyo, Japan).
Zabulica M., Jakobsson T., Ravaioli F., Vosough M., Gramignoli R., Ellis E., Rooyackers O, & Strom S.C. (2021). Gene Editing Correction of a Urea Cycle Defect in Organoid Stem Cell Derived Hepatocyte-like Cells. International Journal of Molecular Sciences, 22(3), 1217.
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2

Generating Human iPSCs from PBMCs

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Peripheral blood mononuclear cells (PBMCs) were reprogrammed to human iPSCs (hiPSCs) using the CytoTune-iPS 2.0 Sendai Reprogramming Kit (ThermoFisher) according to manufacturer’s instructions. All hiPSC lines used in this assay were rigorously tested for pluripotency markers, tested negative for mycoplasma, underwent WGS and underwent short tandem repeat (STR) profiling analysis (LabCorp) to authenticate purity of cell lines. In addition, all hiPSC lines were tested for live alkaline phosphatase activity using the Alkaline Phosphatase Live Stain kit (Thermofisher) according to manufacturer’s instructions.
Fair S.R., Schwind W., Julian D.L., Biel A., Guo G., Rutherford R., Ramadesikan S., Westfall J., Miller K.E., Kararoudi M.N., Hickey S.E., Mosher T.M., McBride K.L., Neinast R., Fitch J., Lee D.A., White P., Wilson R.K., Bedrosian T.A., Koboldt D.C, & Hester M.E. (2022). Cerebral organoids containing an AUTS2 missense variant model microcephaly. Brain, 146(1), 387-404.
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3

Alkaline Phosphatase and SSEA-4 Detection

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Alkaline Phosphatase detection was executed using the Alkaline Phosphatase Live Stain kit (Life Technologies, Cat. # A14353) according to manufacturer’s instructions. Cells were visualized using a Keyence BZ-X700 series microscope. SSEA-4 detection was carried out using the StainAlive SSEA-4 mouse anti-human antibody (Reprocell, Cat. # 09–0097). Antibody was diluted 1:200 in mTeSR1 and cells were incubated in antibody containing medium for 30 min in standard culture conditions. The staining medium was removed and the cells washed gently 2 times with mTeSR1. Fresh media was added and the cells were visualized using a Keyence BZ-X700 series microscope.
Kuang Y.L., Munoz A., Nalula G., Santostefano K.E., Sanghez V., Sanchez G., Terada N., Mattis A.N., Iacovino M., Iribarren C., Krauss R.M, & Medina M.W. (2019). Evaluation of commonly used ectoderm markers in iPSC trilineage differentiation. Stem cell research, 37, 101434.
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4

Directed Differentiation of Pluripotent Stem Cells

Cited 1 time
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hESC lines H1 and H9 were obtained from WiCell Research Institute. iPS-MSC25 (link) (derived from human mesenchymal stem cells) and iPS-Fib225 (link) (derived from human dermal fibroblasts) were a kind gift from George Q. Daley at Children’s Hospital Boston. Essential 8 medium (E8)5 (link), 0.5 mM EDTA, Accutase, ProLong® Antifade reagents, LIVE/DEAD® Cell Viability staining kit, Click-iT® EdU Alexa Fluor® 594 Imaging Kit and Alkaline Phosphatase Live Stain kit were obtained from Life Technologies. Small molecules Y-27632 (ROCK inhibitor, or RI), SB431542, and LDN193189 were from Selleckchem. Matrigel was obtained from BD Biosciences. PNIPAAm-PEG polymer (Mebiol Gel) was from Cosmo Bio, USA. SCID Beige mice were from Charles River Laboratory. Finally, the following antibodies and dilutions were used: Oct4 and Nanog (Santa Cruz Biotech, 1:100); FOXA2 or HNF3β (Santa Cruz Biotech, 1:200); Nestin (Millipore, 1:200); αSMA (Abcom, 1:200).
Lei Y., Jeong D., Xiao J, & Schaffer D.V. (2014). Developing Defined and Scalable 3D Culture Systems for Culturing Human Pluripotent Stem Cells at High Densities. Cellular and molecular bioengineering, 7(2), 172-183.
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5

Alkaline Phosphatase and SSEA-4 Detection

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Alkaline Phosphatase detection was executed using the Alkaline Phosphatase Live Stain kit (Life Technologies, Cat. # A14353) according to manufacturer’s instructions. Cells were visualized using a Keyence BZ-X700 series microscope. SSEA-4 detection was carried out using the StainAlive SSEA-4 mouse anti-human antibody (Reprocell, Cat. # 09–0097). Antibody was diluted 1:200 in mTeSR1 and cells were incubated in antibody containing medium for 30 min in standard culture conditions. The staining medium was removed and the cells washed gently 2 times with mTeSR1. Fresh media was added and the cells were visualized using a Keyence BZ-X700 series microscope.
Kuang Y.L., Munoz A., Nalula G., Santostefano K.E., Sanghez V., Sanchez G., Terada N., Mattis A.N., Iacovino M., Iribarren C., Krauss R.M, & Medina M.W. (2019). Evaluation of commonly used ectoderm markers in iPSC trilineage differentiation. Stem cell research, 37, 101434.
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