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Recombinant full length protein corresponding to Human Interferon alpha 2b.
Database link: P01563
This product was changed from ascites to tissue culture supernatant on 28/11/2017. Lot numbers higher than GR314306-1 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
Our Abpromise guarantee covers the use of ab9386 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.
This antibody can be used in a sandwich ELISA for the determination of human IFN-a2b. It recognizes different epitopes than anti-Interferon alpha 2b antibody [4E10] (HRP) (ab5258). We recommend using this antibody as a capture antibody and ab5258 as a detection antibody.
|Inhibition Assay||1/400 - 1/1000.|
|WB||1/1000 - 1/5000.
At 5 µg/ml, this antibody inhibits 90% of IFN-a2b antiviral activity in vitro: The bioassay was performed using L41 cell line and mouse encephalomyocarditis virus (EMCV). Interferon was pre-incubated with the ab9386 for 2 hours at RT and then added to the cell monolayer. 24 hours later the cells were infected with EMCV and cultured for further 24 hours. Antiviral activity of IFN was evaluated using crystal-violet staining. PAbs to respective interferons and an irrelevant MAb of same isotype were used as controls.
|Indirect ELISA||1/1000 - 1/10000.|
The nitrocellulose membrane was blocked for 1 hour at room temperature with 5% milk in TBST buffer. The individual blots were probed to detect the appropriate model protein for 1 hour at room temperature on a shaker.
ab9386 was used at 1 μg/ml diluted in TBST. After briefly washing the blot membranes with TBST, the secondary antibody was added for 1 hour at room temperature with shaking. The secondary goat anti-mouse AP conjugated antibody was diluted 1:10,000 in TBST.
For full details see figure 11.
Standard curve for the determination of human interferon-alpha 2b by two-epitope ELISA: capture antibody 9D3 (ab9386), peroxidase-labeled antibody 4E10 (ab9388).
Neutralizing activity of monoclonal antibody 9D3 in vitro. Interferon antiviral bioassay: L41 cells, mouse encephalomyacarditis virus (EMCV). Antibodies at 5 µg/ml. Control 1 - without EMCV (non-infected cells), control 2 - without interferon, control 3 - irrelevant antibody.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"