The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 19637364
Use 1-2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 70 kDa.
Use at an assay dependent concentration.
Use a concentration of 1 - 2 µg/ml.
Ubiquitin-editing enzyme that contains both ubiquitin ligase and deubiquitinase activities. Essential component of a ubiquitin-editing protein complex, comprising also RNF11, ITCH and TAX1BP1, that ensures the transient nature of inflammatory signaling pathways. Upon TNF stimulation, deubiquitinates 'Lys-63'-polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains. This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NF-kappa-B. In vitro able to deubiquitinate both 'Lys-48'- and 'Lys-63' polyubiquitin chains. Inhibitor of programmed cell death. Has a role in the function of the lymphoid system.
Belongs to the peptidase C64 family. Contains 7 A20-type zinc fingers. Contains 1 OTU domain.
The A20-type zinc fingers mediate the ubiquitin ligase activity. The OTU domain mediates the deubiquitinase activity.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human placenta tissue labelling TNFAIP3 with ab13597 at 5µg/ml. Staining was enhanced by boiling tissue sections in 10mM sodium citrate buffer, pH6.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes.
Western blot - Anti-TNFAIP3 [59A426] antibody (ab13597)
Overlay histogram showing HepG2 cells stained with ab13597 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13597, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HepG2 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.