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MB Sample ID: SA333683
| Local Sample ID: | DS1-095-013L+ |
| Subject ID: | SU003223 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
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Collection:
| Collection ID: | CO003216 |
| Collection Summary: | For flow cytometry, cells were fixed (1% paraformaldehyde, 0.0075% glutaraldehyde) to prevent antibody agglutination, except for assays using GLUT1.RBD. Samples were labelled with primary antibodies in PBSAG (PBS with 1% (w/v) Glucose and 0.5%(w/v) |Bovine Serum Albumin (BSA)) supplemented with extra 1%(w/v) BSA for 30min, in the dark. METAFORA’s GLUT1.RBD was incubated at 37°C and all remaining antibodies incubated at 4°C. Cells were washed 2x PBSAG and, if required, incubated with appropriate secondary antibody under the same conditions as described for primary. Cells were washed 2x with PBSAG and analysed on a Miltenyi MACSQuant 10 flow cytometer. Data was analysed using FlowJo v10.7 (FlowJo). Reticulocytes were identified by gating on the Hoechst-negative population. 4 Cells were sorted using a BD Influx Cell Sorter (BD Biosciences). BEL-A CRISPR edited populations were single cell sorted based on viability (DRAQ7 negativity). Primary GLUT1 KO cultures were sorted on days 6 or 7 of differentiation using DRAQ7 and the eGFP-fused GLUT1.RBD to purify the negative population with a gate based on non-targeting guide control cells. |
| Sample Type: | Cultured cells |
