H9 hESCs with a normal 46, XX karyotype were acquired from the WiCell Research Institute. The H9 cells were cultured in feeder or feeder-free conditions, depending on the experimental procedure, as described elsewhere2 (link). MDS-L cells were maintained in RPMI-1640 medium (Thermo Fisher Scientific) supplemented with 10% FBS, 1% penicillin–streptomycin and 15 ng ml−1 recombinant human interleukin (IL)-3 (Peprotech). HEK293T cells were purchased from the American Type Culture Collection and maintained in DMEM medium (Thermo Fisher Scientific) with 10% FBS and 1% penicillin–streptomycin. All cells were cultured at 37 °C with 5% CO2 and routinely tested for mycoplasma infection (Universal Mycoplasma Detection Kit, American Type Culture Collection).
Recombinant human interleukin il 3
Manufactured by Thermo Fisher Scientific
Recombinant human interleukin (IL)-3 is a cytokine that plays a role in the regulation of hematopoiesis. It stimulates the proliferation and differentiation of hematopoietic progenitor cells.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Universal mycoplasma detection kit, by American Type Culture Collection (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions)
Fixation buffer, by BioLegend (1 mentions)
Mouse anti human ige, by BD (1 mentions)
Pe mouse anti human cd63, by BD (1 mentions)
2 protocols using recombinant human interleukin il 3
1
Cell Culture Protocols for Pluripotent and Leukemia Cells
2
Multiparametric Basophil Activation Assay
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The following reagents were used for flow cytometry and microscopy staining; Avidin Alexa Fluor TM 488 (ThermoFisher Scientific), APC mouse anti-human CD203c (Clone NP4D6, BD Biosciences), PE mouse anti-human CD63 (Clone H5C6, BD Pharmingen TM ), Alexa Fluor TM 647 mouse anti-human CD63 (Clone H5C6, BD Pharmingen TM ), mouse anti-human IgE labeled with Alexa Fluor 405 NHS Ester (Clone GE-1, Sigma Aldrich GmBH) using the procedure described by Life Technologies.
The reagents used for stimulation of basophils were mouse anti-human IgE (Clone G7-18, BD Pharmingen TM ), recombinant human interleukin (IL)-3 (Peprotech), N-Formyl-Met-Leu-Phe (fMLP, Merck), recombinant major birch pollen allergen (rBet v 1, Biomay).
Buffers used were activation buffer ((Hank's balanced salt solution (Invitrogen) with 20 mM HEPES and 7.5% NaHCO3, pH 7.4)), BD FACS TM lysing solution 10x concentrate (BD Biosciences) and fixation buffer (Biolegend).
The reagents used for stimulation of basophils were mouse anti-human IgE (Clone G7-18, BD Pharmingen TM ), recombinant human interleukin (IL)-3 (Peprotech), N-Formyl-Met-Leu-Phe (fMLP, Merck), recombinant major birch pollen allergen (rBet v 1, Biomay).
Buffers used were activation buffer ((Hank's balanced salt solution (Invitrogen) with 20 mM HEPES and 7.5% NaHCO3, pH 7.4)), BD FACS TM lysing solution 10x concentrate (BD Biosciences) and fixation buffer (Biolegend).
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