Anti-Sup35p-NM Antibody, clone 4A5

Eukaryotic peptide chain release factor GTP-binding subunit (UniProt: P05453; also known as ERF-3, ERF3, ERF2, G1 to S phase transition protein 1, Omnipotent suppressor protein 2, PSI no more protein 2, Polypeptide release factor 3, Translation release factor 3) is encoded by the SUP35 (also known as GST1, PNM2, SAL3, SUF12, SUP2, YD9395.05) gene (Gene ID: 851752) in Saccharomyces cerevisiae (yeast). Sup35 is a GTPase component of the eRF1-eRF3-GTP ternary complex, a complex that mediates translation termination in response to the termination codons UAA, UAG, and UGA. The eRF1-eRF3-GTP complex binds to a stop codon in the ribosomal A-site. The Saccharomyces cerevisiae (yeast) epigenetic element [PSI+] that has multiple structural variants, is a prion of the translation termination factor subunit Sup35p. The yeast Sup35p and the mammalian PrP prion protein share little structural or functional homology except for a repeat region in the amino terminal domain. Sup35 contains three domains: The N-terminal domain (aa 1-123), which is necessary and sufficient for the [PSI+] propagation; the middle domain (M)(aa 124-253) that interacts with the Hsp104 chaperone through the residues 128-148 and is involved in the liquid-liquid phase separation behavior of Sup35 and imparts stability of the prior during mitosis; and the C-terminal domain (aa 254-685) that acts in translation termination. The N-terminal region also contains a prion-forming domain (aa 1-97) that is unstructured in its native soluble form, but forms beta sheets in its amyloid form. Overexpression of the N-domain is shown to increase the frequency of [PSI+] in yeast. Clone 4A5 specifically recognizes the M-domain of Sup35p and is suitable for Western blotting and immunocytochemistry applications. (Ref.: Dergalev, AA., et al. (2019). Int. J. Mol. Sci. 20(11): 2633; Krammer, C., et al. (2009). Proc. Natl. Acad. Sci. USA. 106 (2); 462-467; Krammer, C., et al. (2008). Prion. 2(3); 99-106).

KLH-conjugated linear peptide corresponding to 19 amino acids from the M-domain of SUP35p protein from Saccharomyces cerevisiae (yeast).

Quality Control Testing

Evaluated by Western Blotting in lysate from HEK293 cells transfected with HA-tagged Sup35 NM.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Sup35NM (M Domain) in lysate from HEK293 cells transfected with HA-tagged Sup35NM.

Tested Applications

Immunocytochemistry Analysis: A representative lot detected Sup35/HEFE-ME in Immunocytochemistry applications (Krammer, C., et al. (2009). Proc Natl Acad Sci USA. 106(2):462-7).

Western Blotting Analysis: A representative lot detected Sup35/HEFE-ME in Western Blotting applications (Riemschoss, K., et al. (2019). Life Sci Alliance. 2(4):e201800280).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 4A5 is a rat monoclonal antibody that detects Eukaryotic peptide chain release factor GTP-binding subunit (SUP35p). It targets an epitope within 19 amino acids from the N-terminal half.

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

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommended storage: +2°C to +8°C.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.