Recommended ELISA procedure for human IFN alpha 2b determination: 1. Use 96-well Nunc Maxisorp plates for the assay. Coat microwells with 100 µl/well of antibody 9D3 (ab9386) diluted to 10 µg/ml in coating buffer (0.05 M Na-carbonate, pH 9.5). Incubate overnight at +4°C. 2. Aspirate wells and block with 150 µl/well of blocking buffer (2% BSA in PBS). Incubate at RT for 1 h. 3. Aspirate wells and wash 3 times with PBST (0.1% Tween-20 in PBS). 4. Prepare standard and sample dilutions in PBST. Recommended IFN alpha 2b concentrations: from 100 pg/ml to 100 ng/ml. Add 100 µl of each standard and sample into appropriate wells. Incubate at RT for 2 h. 5. Aspirate and wash 5 times with PBST. 6. Dilute detection antibody 4E10-HRP (ab5258) 1:1000 in PBST. Add 100 µl of diluted detection antibody to each well. Incubate at RT for 1 h. 7. Aspirate and wash 10 times with PBST. 8. Add 100 µl of substrate solution (TMB) to each well. Incubate at RT for 10-20 min. 9. Add 50 µl of stop solution (2 N H2SO4) to each well. Read absorbance at 450 nm.

This product was changed from ascites to tissue culture supernatant on 28/11/2017. Lot numbers higher than GR308991-1 and GR308991-2 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.