FACS-isolated stromal cells were cultured in 10% FBS/DMEM in a 48-well tissue culture plate. When the cells were 80% confluent, old media was removed and cells were incubated in 150 μl 10% FBS/DMEM containing 10 μM purmorphamine (Calbiochem 540220) or DMSO control. In separate experiments, cultured stromal cells were treated with 10% FBS/DMEM containing 5 μg/ml recombinant Ihh protein (R&D 1705-HH-025) in 0.1% BSA or a BSA only control. In separate experiments, cells were transfected with miRNA mimics for miR-26a (Ambion, Assay MC10249) or miR-26b (Ambion, Assay MC12899) or a negative control (Ambion 4464076). After 22 h, media was removed and centrifuged at 10,000g for 5 min. To measure the concentration of Hgf in the media, 10 μl of the supernatant was used in each well of the Mouse/Rat HGF Quantikine ELISA Kit (R&D Systems, MHG00) and the plate processed according to manufacturer’s instructions. Each sample was analyzed in duplicate.
Assay mc12899
Manufactured by Thermo Fisher Scientific
The Assay MC12899 is a laboratory instrument designed for use in analytical and research applications. It is capable of performing various types of assays, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using assay mc12899
1
Regulation of Stromal Cell HGF Secretion
2
Regulation of Stromal Cell HGF Secretion
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FACS-isolated stromal cells were cultured in 10% FBS/DMEM in a 48-well tissue culture plate. When the cells were 80% confluent, old media was removed and cells were incubated in 150 μl 10% FBS/DMEM containing 10 μM purmorphamine (Calbiochem 540220) or DMSO control. In separate experiments, cultured stromal cells were treated with 10% FBS/DMEM containing 5 μg/ml recombinant Ihh protein (R&D 1705-HH-025) in 0.1% BSA or a BSA only control. In separate experiments, cells were transfected with miRNA mimics for miR-26a (Ambion, Assay MC10249) or miR-26b (Ambion, Assay MC12899) or a negative control (Ambion 4464076). After 22 h, media was removed and centrifuged at 10,000g for 5 min. To measure the concentration of Hgf in the media, 10 μl of the supernatant was used in each well of the Mouse/Rat HGF Quantikine ELISA Kit (R&D Systems, MHG00) and the plate processed according to manufacturer’s instructions. Each sample was analyzed in duplicate.
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