Anti-TNF alpha antibody [52B83]

4 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)

  ab1793 staining TNFα in RAW 264.7 cells treated with (R,S)-rolipram ((R,S)-Rolipram, Selective PDE4 inhibitor ab120029), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R,S)-rolipram, as described in literature.
  The cells were incubated at 37°C for 24h in media containing different concentrations of (R,S)-Rolipram, Selective PDE4 inhibitor ab120029 ((R,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 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed 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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  Flow Cytometry (Intracellular) - Anti-TNF alpha antibody [52B83] (ab1793)

  Overlay histogram showing RAW 264.7 cells stained with ab1793 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1793, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in RAW 264.7 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.
  
  Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  

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  Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)

  ab1793 staining TNFα in RAW 264.7 cells treated with (R)-(-)-rolipram ((R)-(-)-Rolipram, Selective PDE4 inhibitor ab120031), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R)-(-)-rolipram , as described in literature.
  The cells were incubated at 37°C for 24h in media containing different concentrations of (R)-(-)-Rolipram, Selective PDE4 inhibitor ab120031 ((R)-(-)--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 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed 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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  Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)

  ab1793 staining TNFα in RAW 264.7 cells treated with (S)-(+)-rolipram ((S)-(+)-Rolipram, PDE4 inhibitor 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 (S)-(+)-Rolipram, PDE4 inhibitor 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 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.