Blastocysts were flushed out from the uterus of Usp16+/− or Usp162lox/+:CAGG-Cre E3.5 female mice with a 27.5 gauge needle connected to 1 ml syringe filled with 1 ml 2i medium45 (link). Blastocysts were then transferred to feeder cell plates with 200 μl sterile tip and cultured with 2i medium46 (link) or regular ESC medium for 5–8 days. ESC-like colonies formed from ICM outgrowths were handpicked using sterile pipette tips and dissociated by 0.25% trypsin and expanded in ESC medium. For proliferation rate assay, 1 × 104 ESCs were seeded in 12 well plates and cell numbers were counted and the same seeding procedure was repeated every other day. Western blot assay and qPCR were performed using standard protocol included in the method section47 (link). Antibodies include anti-Usp16 (1:1000)26 (link), anti-ubH2A (Millipore, 05-678, 1:1000), anti-ubH2B (Millipore, 05-1312, 1:1000), anti-H3 (Abcam, ab100938, 1:5000), anti-GAPDH (Sigma, G9545, 1:5000), Anti-Nanog (R&D Systems, AF2729, 1: 2000), Anti-Oct4 (Santa Cruz Biotechnology, SC9081, 1:2000), and anti-Sox2 (Cell signaling, 3579, 1:2000). Alkaline phosphatase substrate kit was from Vector laboratory (SK-5100).
Ab100938
Manufactured by Merck Group
Ab100938 is a laboratory equipment product offered by Merck Group. It serves as a core tool for various research applications. The product's primary function is to facilitate essential laboratory processes. Further details on its intended use or specific applications are not available.
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Lab products found in correlation
2 protocols using ab100938
1
Isolation and Characterization of Embryonic Stem Cells from Usp16 Mutant Mice
2
Isolation and Characterization of Embryonic Stem Cells from Usp16 Mutant Mice
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Blastocysts were flushed out from the uterus of Usp16+/− or Usp162lox/+:CAGG-Cre E3.5 female mice with a 27.5 gauge needle connected to 1 ml syringe filled with 1 ml 2i medium45 (link). Blastocysts were then transferred to feeder cell plates with 200 μl sterile tip and cultured with 2i medium46 (link) or regular ESC medium for 5–8 days. ESC-like colonies formed from ICM outgrowths were handpicked using sterile pipette tips and dissociated by 0.25% trypsin and expanded in ESC medium. For proliferation rate assay, 1 × 104 ESCs were seeded in 12 well plates and cell numbers were counted and the same seeding procedure was repeated every other day. Western blot assay and qPCR were performed using standard protocol included in the method section47 (link). Antibodies include anti-Usp16 (1:1000)26 (link), anti-ubH2A (Millipore, 05-678, 1:1000), anti-ubH2B (Millipore, 05-1312, 1:1000), anti-H3 (Abcam, ab100938, 1:5000), anti-GAPDH (Sigma, G9545, 1:5000), Anti-Nanog (R&D Systems, AF2729, 1: 2000), Anti-Oct4 (Santa Cruz Biotechnology, SC9081, 1:2000), and anti-Sox2 (Cell signaling, 3579, 1:2000). Alkaline phosphatase substrate kit was from Vector laboratory (SK-5100).
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