PMC:3724992 / 3948-5071 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3724992","sourcedb":"PMC","sourceid":"3724992","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3724992","text":"Peptide-binding assay\nTo determine whether the candidate epitopes can bind to HLA-A*0201 molecules, up-regulation of peptide-induced HLA-A*0201 molecules on T2 cells was examined. Briefly, according to previous reference, 1 × 106 T2 cells were incubated with 50 μM of the synthesized peptides in serum-free RPMI 1640 medium supplemented with β2-microglobulin (Sigma) at a concentration of 3 μg/ml for 16 h at 37 °C, 5 % CO2 [18]. Expression of HLA-A*0201 on T2 cells was then determined with the FACS Calibur flow cytometer (Becton–Dickinson, USA), by staining with primary anti-HLA-A2 Ab derived from BB7.2 and FITC-labeled goat-antimouse IgG (BD Biosciences Pharmingen, USA) secondary antibody. The data were analyzed using Cell Quest software (Becton–Dickinson, USA). The Fluorescence index (FI) was calculated as follows: FI = (mean FITC fluorescence with the given peptide − mean FITC fluorescence without peptide)/(mean FITC fluorescence without peptide). Samples were measured in triplicate and then mean FI was calculated. An octapeptide mHpa (519–526) (FSYGFFVI) derived from mouse Hpa was served negative control.","divisions":[{"label":"title","span":{"begin":0,"end":21}}],"tracks":[{"project":"2_test","denotations":[{"id":"23754640-16534571-63236739","span":{"begin":425,"end":427},"obj":"16534571"}],"attributes":[{"subj":"23754640-16534571-63236739","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93ec9c","default":true}]}]}}