TNFAIP3 Mouse Monoclonal Antibody [Clone ID: 59A426]

CAT#: TA336834

Mouse Monoclonal A20/TNFAIP3 Antibody (59A426)

Product Images

Simple Western: A20/TNFAIP3 Antibody (59A426) TA336834 - Lane view shows a specific band for A20/TNFAIP3 in 0.05 mg/ml of Jurkat lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

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  Immunohistochemistry-Paraffin: A20/TNFAIP3 Antibody (59A426) TA336834 - Human placenta probed with A20 antibody at 5 ug/ml, cytoplasmic staining of decidual cells is observed. Human tissue TMA was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

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  Flow Cytometry: A20/TNFAIP3 Antibody (59A426) TA336834 - An intracellular stain was performed on RH-30 cells with A20/TNFAIP3 Antibody (59A426) TA336834PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin.

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  Immunocytochemistry/Immunofluorescence: A20/TNFAIP3 Antibody (59A426) TA336834 - Caco-2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-A20/TNFAIP3 Antibody [59A426] TA336834 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

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  Flow Cytometry: A20/TNFAIP3 Antibody (59A426) TA336834 - An intracellular stain was performed on SK-MEL-28 cells with A20/TNFAIP3 (59A426) antibody TA336834AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

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  Immunocytochemistry/Immunofluorescence: A20/TNFAIP3 Antibody (59A426) TA336834 - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-A20/TNFAIP3 (59A426) at 10 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective

• Western Blot: A20/TNFAIP3 Antibody (59A426) TA336834 - Human Jurkat lysate probed with A20 antibody at 4 ug/ml.

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Specifications

Product Data

• Clone Name: 59A426
• Applications: FC, IHC, WB
• Recommended Dilution: Flow Cytometry: 2.5 ug/ml, Immunohistochemistry-Paraffin: 5 - 10 ug/ml, Flow (Intracellular): 2.5 ug/ml, Western Blot: 2-4 ug/ml, Immunoprecipitation: 1-2 ug/ml, Immunocytochemistry/ Immunofluorescence: 10 ug/ml, Immunohistochemistry: 5 - 10 ug/ml, Simple Western: 1:100, CyTOF-ready
• Reactivities: Human, Mouse, Rat
• Host: Mouse
• Isotype: IgG1, kappa
• Clonality: Monoclonal
• Immunogen: Full length recombinant human A20. The epitope has been mapped to the C-terminal portion of A20 (amino acids 440-790)
• Formulation: PBS containing 0.05% BSA, 0.05% Sodium Azide. Store at -20C. Avoid freeze-thaw cycles.
• Concentration: lot specific
• Purification: Protein G purified
• Conjugation: Unconjugated
• Storage: Store at -20°C as received.
• Stability: Stable for 12 months from date of receipt.
• Gene Name: TNF alpha induced protein 3
• Database Link:
  NP_006281
  Entrez Gene 21929 MouseEntrez Gene 683206 RatEntrez Gene 7128 Human
  UniProt ID P21580
• Background: A20 (TNFAIP3) is an important regulator of proinflammatory signaling pathways, including NK-kB sginaling. A20 is upregulated through NF-kB (p50/p55) activation, and subsequently down regulates NF-kB through its dual function as a deubiquitinase and ubiquitin ligase (Evans, 2005). The A20 monoclonal antibody (clone 59A426) has been instrumental in discovering various nuances of NF-kB regulation and key to defining A20 biology.
• Synonyms: A20; OTUD7C; TNFA1P2
• Note: The A20 antibody (clone 59A426) has also been used in Immunohistochemistry-Paraffin (Metellus, 2010) and Flow Cytometry (intracellular) (Hjelmeland, 2010). Multiple A20 cleavage fragments have been described in Western Blot, see Coornaert (2008) and Hailfinger (2009) for additional details.

Reference Data

• Protein Families: Druggable Genome
• Protein Pathways: NOD-like receptor signaling pathway