title: aCD3_CD19_HD_14
organism: Homo sapiens
description: Protocols: Lithium Heparin (LiHep)-anticoagulated blood was withdrawn from a total of 24 healthy donors, 25 SLE patients and 23 pSS patients. PBMCs were isolated from LiHep-anticoagulated blood. The collected mononuclear cells were washed twice with PBS. cells were rested for 30 min at 37°C in ImmunoCult-XF T cell expansion medium (Stemcell). 96-well plate were coated with 0.1 µg/ml anti-CD3 (OKT3, eBioscience) for 2h at 37°C or PBS as control, washed two times with PBS, blocked with PBS/1% BSA for 30 min and washed two times with PBS. 1x106 PBMCs were seeded per well and incubated at 37°C and 5% CO2 in a humidified incubator for 18hours. The PicoPure RNA Kit (Thermo Fisher) was used for purification of total RNA from cells. Library generation has been completed using the SMARTer cDNA Synthesis and Nextera XT chemistry. With 500pg total RNA, cDNA pools were prepared using the SMART-Seq v4 Kit (Clontech Laboratories) to generate full-length cDNA from total RNA.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-01-31
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-01-31
attributes.tag: External Id
attributes.value: SAMEA111385102
attributes.tag: INSDC center alias
attributes.value: Systems Biology of Inflammation, the German Rheumatism Research Centre Berlin, a Leibniz Institute (DRFZ)
attributes.tag: INSDC center name
attributes.value: Systems Biology of Inflammation, the German Rheumatism Research Centre Berlin, a Leibniz Institute (DRFZ)
attributes.tag: INSDC first public
attributes.value: 2023-01-31T00:19:52Z
attributes.tag: INSDC last update
attributes.value: 2023-01-31T00:19:52Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12203:aCD3_CD19_HD_14
attributes.tag: age
attributes.value: 29
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: B cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: normal
attributes.tag: immunophenotype
attributes.value: CD19+
attributes.tag: individual
attributes.value: HD_14
attributes.tag: organism part
attributes.value: blood
attributes.tag: sample name
attributes.value: E-MTAB-12203:aCD3_CD19_HD_14
attributes.tag: sex
attributes.value: female