Product nameAnti-TNFAIP3 antibody [59A426]
See all TNFAIP3 primary antibodies
DescriptionMouse monoclonal [59A426] to TNFAIP3
Tested applicationsSuitable for: IHC-P, Flow Cyt, WB, ICC/IF, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Recombinant fusion protein of full length TNFAIP3 (Human).
EpitopeThe epitope has been mapped to the C-terminal portion of A20, amino acids 440-790.
- WB: Daudi and HeLa cell lysates.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium azide
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab13597 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: 19637364|
|Flow Cyt||Use 1-2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 70 kDa.|
|ICC/IF||Use at an assay dependent concentration.|
|IP||Use a concentration of 1 - 2 µg/ml.|
FunctionUbiquitin-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.
Sequence similaritiesBelongs to the peptidase C64 family.
Contains 7 A20-type zinc fingers.
Contains 1 OTU domain.
DomainThe A20-type zinc fingers mediate the ubiquitin ligase activity.
The OTU domain mediates the deubiquitinase activity.
Cellular localizationCytoplasm. Nucleus.
- Information by UniProt
- A20 antibody
- AISBL antibody
- MGC104522 antibody
All lanes : Anti-TNFAIP3 antibody [59A426] (ab13597) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : TNFAIP3 knockout HeLa cell lysate
Lane 3 : Jurkat cell treated with 5ng/ml PMA for 48 hours and then treated with 2µg/ml PHA for 48 hours, whole cell lysate
Lane 4 : Untreated Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 80 kDa why is the actual band size different from the predicted?
Lanes 1-4: Merged signal (red and green). Green – ab13597 observed at 80 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab13597 was shown to react with TNFAIP3 in wild-type HeLa cells in Western blot. Loss of signal was observed when knockout sample ab257112 was used. Wild-type and TNFAIP3 knockout samples were subjected to SDS-PAGE. ab13597 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Anti-TNFAIP3 antibody [59A426] (ab13597) at 4 µg/ml + Jurkat cell lysate
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.
All lanes : Anti-TNFAIP3 antibody [59A426] (ab13597) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 90 kDa why is the actual band size different from the predicted?
Additional bands at: 15 kDa, 34 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
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.
This product has been referenced in:
- Yang M et al. Decreased A20 expression on circulating CD56bright NK cells contributes to a worse disease status in patients with ankylosing spondylitis. Clin Exp Immunol N/A:N/A (2019). Read more (PubMed: 31206174) »
- Lu J et al. Melatonin Suppresses Microglial Necroptosis by Regulating Deubiquitinating Enzyme A20 After Intracerebral Hemorrhage. Front Immunol 10:1360 (2019). Read more (PubMed: 31258534) »