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Full length native protein (purified) (Human).
Our Abpromise guarantee covers the use of ab1793 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||1/10. (methanol fixed cells)
Fixed in acetone for 10 minutes
|ICC/IF||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 17 kDa.
TNF-alpha is normally secreted as a homotrimer with a molecular mass of 52 kDa. Monomeric TNF-alpha (17.4 kDa) is not biologically active.
A reduced sample treatment and 15% SDS-Page was used.
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab1793 staining TNFα in RAW 264.7 cells treated with (S)-(+)-rolipram (ab120030), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (S)-(+)-rolipram , as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120030 ((S)-(+)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793(5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
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