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Our Abpromise guarantee covers the use of ab3523 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. PubMed: 28186091|
|WB||1/200 - 1/800. Detects a band of approximately 135 kDa.
Abcam recommends using 5% BSA as the blocking agent.
Note: Regarding using this antibody in WB on mouse lung tissue: "However, iNOS protein was below the limit of detection following any level of hypoxic exposure (positive control was lipopolysaccharide-treated mouse spleen) and using several different antibodies" (from PubMed ID 11686901)
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab3523 overnight at 4°C. Antibody binding was detected using Goat anti Rabbit IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx
The cells were blocked with 5% milk for 2 hours at 22°C.
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