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Recombinant full length protein (Human).
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.
Our Abpromise guarantee covers the use of ab5258 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"