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Ab0300

Manufactured by Merck Group
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The AB0300 is a laboratory equipment product manufactured by Merck Group. It is a precision instrument designed for scientific research and analysis. The core function of the AB0300 is to perform accurate measurements and data collection. Further details about the intended use or specific applications of this product are not available.

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Lab products found in correlation

A21422, by Thermo Fisher Scientific (2 mentions) Mab430, by Thermo Fisher Scientific (2 mentions)

4 protocols using ab0300

1

Alkaline Phosphatase and Immunofluorescence

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Alkaline phosphatase staining was performed using a kit (Sigma, AB0300) according to the manufacturers protocol. Cells were imaged under a Zeiss microscope at 20X objective. Immunofluorescence experiments were performed according to Whyte et al., 2012 (31 (link)). Antibodies used for immunofluorescence include: SSEA-1 (Millipore, MAB430) 1:2000 and AlexaFluor 555 nm (Life Technologies, A21422) 1:1000. Brightfield imaging was done with a Nikon Ts microscope with a 20X objective.
Petell C.J., Alabdi L., He M., San Miguel P., Rose R, & Gowher H. (2016). An epigenetic switch regulates de novo DNA methylation at a subset of pluripotency gene enhancers during embryonic stem cell differentiation. Nucleic Acids Research, 44(16), 7605-7617.
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2

Embryoid Body Characterization via Staining

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Bright field images of embryoid bodies (EBs) were obtained with Zeiss microscope using a 10X objective. Alkaline phosphatase staining was performed using solutions supplied by an alkaline phosphatase staining kit (Sigma, AB0300). Cells were cross-linked with 1% formaldehyde for 5 min, followed by quenching with a final concentration of 150 mM glycine. Cells were washed twice with 1xPBS, then twice with combined staining solution (BCIP and NBT). The stain was developed in the dark for 5 min, then washed three times with 1XPBS. SSEA-1 immunofluorescence was performed using the following antibodies: anti-SSEA-1 (Millipore, MAB430) and AlexaFluor 555 nm (Life Technologies, A21422). SSEA-1 and Alkaline phosphatase staining were imaged using 20X objectives under Nikon Ts and Zeiss microscopes, respectively.
AlAbdi L., Saha D., He M., Dar M.S., Utturkar S.M., Sudyanti P.A., McCune S., Spears B.H., Breedlove J.A., Lanman N.A, & Gowher H. (2020). Oct4-Mediated Inhibition of Lsd1 Activity Promotes the Active and Primed State of Pluripotency Enhancers. Cell reports, 30(5), 1478-1490.e6.
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3

Alkaline Phosphatase Activity in iPSCs

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AP activity was studied in the whole 10-cm Petri dishes (Costar Corning Incorporated) with newly appeared colonies from the two patients using the alkaline phosphatase blue membrane substrate solution kit (AB0300, Sigma-Aldrich Química S.A.) according to the manufacturer’s guidelines. Furthermore, iPSC clones with more than 20 passages were seeded onto a 6 well feeder plate (Costar Corning Incorporated). When colonies appeared, AP activity was also determined. All samples were observed in a Nikon stereomicroscope SMZ745 coupled to a Nikon Digital Slight DS-Fi2 camera (Nikon).
Castro-Viñuelas R., Sanjurjo-Rodríguez C., Piñeiro-Ramil M., Hermida-Gómez T., Rodríguez-Fernández S., Oreiro N., de Toro J., Fuentes I., Blanco F.J, & Díaz-Prado S. (2020). Generation and characterization of human induced pluripotent stem cells (iPSCs) from hand osteoarthritis patient-derived fibroblasts. Scientific Reports, 10, 4272.
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4

Quantitative Alkaline Phosphatase Staining

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Five-thousand serum LIF cultured cells were seeded, fixed after 5 d (4% formaldehyde), and stained with alkaline phosphatase blue membrane substrate solution (Sigma-Aldrich AB0300), according to the manufacturer's protocol.
Ignatova V.V., Stolz P., Kaiser S., Gustafsson T.H., Lastres P.R., Sanz-Moreno A., Cho Y.L., Amarie O.V., Aguilar-Pimentel A., Klein-Rodewald T., Calzada-Wack J., Becker L., Marschall S., Kraiger M., Garrett L., Seisenberger C., Hölter S.M., Borland K., Van De Logt E., Jansen P.W., Baltissen M.P., Valenta M., Vermeulen M., Wurst W., Gailus-Durner V., Fuchs H., Hrabe de Angelis M., Rando O.J., Kellner S.M., Bultmann S, & Schneider R. (2020). The rRNA m6A methyltransferase METTL5 is involved in pluripotency and developmental programs. Genes & Development, 34(9-10), 715-729.
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