GEMM-CFU were assayed using a complete growth media/methylcellulose (Sigma-Aldrich) based colony assay (Cortdy 1995 , Seed et al. 2002a ). The complete medium was comprised of: methylcellulose (Sigma-Aldrich) in Iscove’s MDM (Life Technologies), HI-FBS, bovine serum albumin (Sigma-Aldrich), bovine pancreatic insulin (Sigma-Aldrich), human transferrin (Sigma-Aldrich), 2-mercaptoethanol (Sigma- Aldrich), L-glutamine (Life Technologies), recombinant stem cell factor (rSCF; Pharmingen), recombinant interleukin-3 (rIL-3; R&D Systems, Minneapolis, MN), recombinant human interleukin-6 (rhIL-6; R&D Systems), and recombinant human erythropoietin (rh erythropoietin, R&D Systems). Nucleated bone marrow cells were diluted to 1.5 × 105 cells/ml in IMDM with 2% HI-FBS; 0.3 ml of diluted cells were mixed with 3.0 ml of the complete medium. Cell/media mix (1 ml) was dispensed to each 35 mm plate. GEMM-CFU colonies showing all four lineages were scored 14 days after incubation in a 37°C humidified environment containing 5% CO2.
Ril 3
Manufactured by R&D Systems
RIL-3 is a laboratory instrument designed for rapid imaging and analysis of western blot and other gel-based assays. The core function of RIL-3 is to capture high-resolution digital images of electrophoretically separated proteins or nucleic acids and provide quantitative analysis of band intensity and molecular weight.
Automatically generated - may contain errors
Lab products found in correlation
Bovine pancreatic insulin, by Merck Group (1 mentions)
Human transferrin, by Merck Group (1 mentions)
Hi fbs, by Merck Group (1 mentions)
Iscove s mdm, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
2 protocols using ril 3
1
GEMM-CFU Assay for Bone Marrow Cells
2
Basophil Activation by IL-3 and T-DC Supernatant
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Basophils (1 Â 10 6 /mL/well) were plated in 96-well plates and cultured for 16 hours in IMDM 10% FBS AE the following stimuli: 5 ng/mL rIL3 (R&D Systems) and supernatants from T cells cocultured with DCs (containing 60-200 pg/mL IL3). IL3-neutralizing or isotype control antibodies (R&D Systems) were added at 1 mg/mL. Basophil activation was measured by FACS analysis by quantitating CD203c expression, as reported in refs. 26, 27.
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