characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2013-10-26T17:01:37Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:46:51Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2240927
characteristics.characteristic: External Id
characteristics.text: Swiss Federal Institute of Technology Zurich
characteristics.characteristic: INSDC center name
characteristics.text: 2013-10-26T17:01:37Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:46:51Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS351324
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1896:one mouse
characteristics.characteristic: Submitter Id
characteristics.text: 8 week
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000034
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Immunised with NP-CGG (chicken gamma globulin (CGG) conjugated to 4-hydroxy-3-nitrophenylacetyl)
characteristics.characteristic: clinical information
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: All animal experiments were performed under the guidelines and protocols approved by the Basel-Stadt cantonal veterinary office (Basel-Stadt Kantonales VeterinŠramt Tierversuchsbewilligung #2582). Female BALB/c mice (n=10, Charles Rivers Laboratories) 6 to 8 weeks old were housed under specific pathogen-free conditions and were maintained on a normal chow diet. Single cell suspensions of total splenocytes and bone marrow were obtained as described previously 4. Briefly, tibiae, femora, and spleens were collected into RPMI plus 10 % fetal bovine serum (FBS, Sigma). Bone marrow cells were flushed out using a 26-gauge syringe (Braun) and spleens were disintegrated using syringe and forceps. Cells were filtered through a 70-uM cell strainer (BD) into 15 mL falcon tubes (Falcon, BD) and subsequently centrifuged at 1500 rpm for 10 min at 4 degrees C. Red blood cells were lysed for 3 min in red blood cell lysis buffer (eBioscience). Cell suspension was then washed once with 10 mL RPMI + 10% FBS. Finally, cells were resuspended in 1 mL PBS + 0.5 % bovine serum albumin (BSA) + 2 mM ethylenediaminetetracetic acid (EDTA, buffer 1). Bone marrow cells were depleted of CD45R-positive cells before plasma cell isolation by incubation with anti-CD45R-biotin antibody (eBioscience, 13-0452) for 15 min on ice. After washing the cell suspension twice in BSA-free PBS supplemented with 2 mM EDTA (buffer 2), 30 ul of washed streptavidin-coupled Dynabeads (M-280, Invitrogen) was added and incubated for 20 min on ice. Following magnetic selection, the negative fraction was used in the subsequent steps. For plasma cell enrichment, spleen and CD45R-depleted bone marrow cell fractions were incubated with anti-CD138-biotin antibody (BD, 553713) for 15 min on ice. Following two wash steps in buffer 2, 30 ul of Dynabeads were added and incubated for 20 min on ice as before. Bead-bound cells were manually counted on a Neubauer cell counter, buffer removed by separation on the magnet, and beads plus cells were lysed in 1 mL of TRIzol¨ reagent (Invitrogen) and stored at -80 degrees C until further usage. Following cell isolation, isolated spleen and bone marrow cells were combined from nine mice ('9M', ~2.5 x 107 ASCs) and one mouse ('1M', ~3 x 106 ASCs) samples.
characteristics.characteristic: description
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: spleen and bone marrow
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: Balb/c
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000602
characteristics.characteristic: strain
characteristics.text: one mouse
characteristics.characteristic: title