Anti-SET (I2PP2A) Antibody, N-term Antibody, clone 10E7

Protein SET (UniProt Q01105; also known as HLA-DR-associated protein II, IGAAD, Inhibitor of granzyme A-activated DNase, PHAPII, I-2PP2A, Phosphatase 2A inhibitor I2PP2A, TAF-I, Template-activating factor I) is encoded by the SET gene (Gene ID 6418) in human. Protein SET (patient SE translocation ) adopted its name when it was originally identified in a leukemia study in 1992. SET interacts with signaling pathways that promote normal cell growth and proliferation, as well as tumorigenesis and metastasis. SET protein inhibits PP2A by enhancing lyase activity of P450c17, a substrate for PP2A when it is Ser- and Thr-phosphorylated. Alternative splicings result in multiple SET isoforms, including SET-alpha (TAF-I alpha), SET-beta (TAF-I beta), where the N-terminal 37-amino acid sequence in Set-alpha is replaced with a different sequence of a shorter length in Set-beta. In addition, there exists a shorter spliced Set- isoform without the nuclear localization signal (NLS). Both Set-alpha and Set-beta are predominantly localized in the nucleus, but differ in their expression levels among various cell and tissue types. Set-beta is predominant in the brain relative to Set-alpha, whereas the opposite pattern was observed in tissues such as the spleen. Set-beta functions have been described both in the nucleus and cytoplasm. The presence of a nuclear localization signal (NLS) and phosphorylation govern Set- isoforms cellular localization. Proteolytic cleavage of Set-beta at Lys176 by granzyme A or at Asn-175 by asparagine endopeptidase (AEP) results in the release of GzmA-activated DNase (GAAD) from a multiprotein complex. Uninhibited in the absence of the full-length Set-eta, GAAD causes cell death by its DNA-nicking activity.

Clone 10E7 detected both full-length human SET (I2PP2A) and the N-terminal fragment I2NTF generated by Asn endopeptidase (AEP) cleavage (Basurto-Islas, G., et al. (2013). J. Biol. Chem. 288(24):17495-17507).

Recombinant human SET (I2PP2A).

Anti-SET (I2PP2A), N-term, clone 10E7, Cat. No. MABN1843, is a highly specific mouse monoclonal antibody that targets I2PP2A/SET and has been tested in Western Blotting.

Research Category
Neuroscience

Western Blotting Analysis: 0.5 µg/mL from a representative lot detected SET (I2PP2A) isoforms in 10 µg of HEK293 and HeLa cell lysates.

Western Blotting Analysis: A representative lot detected endogenous monkey SET (I2PP2A) as well as full-length human SET constructs (Wild-type, R179A/K180A/R181A, K168A/R169A) exogenously expressed in COS-7 cells (Arif, M., et al. (2014). J. Biol. Chem. 289(40):27677-27691).

Western Blotting Analysis: A representative lot detected full-length Myc-tagged human SET (I2PP2A) and its N-terminal cleavage product (NCP) in transfected COS-7 cells. Only NCP and Asn endopeptidase-/AEP-cleaved N-terminal fragment I2NTF, but not the full-length SET, were detected in COS-7 co-transfected with full-length SET and AEP, while acidic pH treatment of the COS-7 transfectants resulted in the detection of only I2NTF, but not NCP (Basurto-Islas, G., et al. (2013). J. Biol. Chem. 288(24):17495-17507).

Western Blotting Analysis: A representative lot detected both endogenous SET (I2PP2A) in untransfected PC12 cells and exogenously overexpressed SET (I2PP2A) in PC12 rat adrenal pheochromocytoma cells.

Evaluated by Western Blotting in Jurkat cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected SET (I2PP2A) isoforms in 10 µg of Jurkat cell lysate.

~33-40 kDa observed. Band size appears larger than the calculated molecular weights of 33.36/31.97/30.86/31.14 kDa (human isoform 1 (SET-alpha)/2 (SET-beta)/3/4; Met1 removed) due to posttranslational modification (acetylation and/or phosphorylation). Uncharacterized bands may be observed in some lysate(s).

Protein G purified.

Format: Purified

Purified mouse IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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