Anti-mS2i6A Antibody, clone CL-3

2-methylthio-N6-isopentenyladenosine (mS2i6A) is a hypermodified modified nucleoside that has been shown to be present in mitochondrial tRNA. It reads codons starting with uridine. The mS2 group of mS2i6A is critical for the accurate decoding of Tyr and Phe codons. Higher concentrations of mS2i6A are found in tissues with a high energy demand, such as muscle and brain. One of the major role of mS2i6A is to stabilize the stacking interactions of the anticodon loop helix and thereby prevent codon misreading. Its deficiency has been linked to a higher rejection rate of tRNA4Leu by more aggressive proofreading on the wild-type ribosome. However, it is shown that mS2i6A has no effect on codon-anticodon interactions on wild-type ribosomes as long as aminoacyl-tRNA is in ternary complex with EF-Tu and GTP. Also, it has been reported that the lack of mS2i6A does not decrease the ability of the tRNA to be charged with its cognate amino acid, but it alters the efficiency with which the tRNA is bound to the ribosome. CDK5RAP1, which inhibits the activity of CDK5, is shown to be an N6-isopentenyladenosine methylthiotransferase. It converts the RNA modification N6-isopentenyladenosine (i6A) in mS2i6A post-synthetically. This conversion is observed both in the mitochondrial as well as nuclear RNA species. (Ref.: Wilson, RK and Roe, BA (1989). Proc. Natl. Acad. Sci. USA. 86(2); 409-413, Diaz, I., and Ehrenberg, M (1991). J. Mol. Biol. 222(4); 1161-71; Reiter, V et al. (2012). Nucleic Acids Res. 40(13); 6235-40).

A synthesized mS2i6A oligonucleotide conjugated to BSA.

Anti-MS2i6A, clone CL-3, Cat. No. MABS1295, is a mouse mouse monoclonal antibody that detects mS2i6A and has been tested for use in Dot Blot, ELISA, Immunocytochemistry, and Immunofluorescence.

ELISA Analysis: A representative lot detected mS2i6A in ms2i6A or i6A wells (Courtesy of Fan-Yan Wei, Ph.D., Faculty of Life Sciences, Kumamoto University Japan).

Immunofluorescence Analysis: A representative lot detected mS2i6A in HeLa cells treated with mitotracker (Courtesy of Fan-Yan Wei, Ph.D., Faculty of Life Sciences, Kumamoto University Japan).

Dot Blot Analysis: A 1:200 dilution from a representative lot detected mS2i6A in Wild type (WT) total RNA and Cdk5rap1 KO mice total RNA samples from liver (Courtesy of Fan-Yan Wei, Ph.D., Faculty of Life Sciences, Kumamoto University Japan).

Clone 3 (CL-3) recognizes wild-type total RNA but not knockout total RNA.

Format: Purified

Evaluated by Immunocytochemistry in HeLa cells with mitotracker.

Immunocytochemistry Analysis: A 1:500 dilution of this antibody detected mS2i6A in HeLa cells with Mitotracker.

Concentration: Please refer to lot specific datasheet.