For research use only. Not for use in diagnostic procedures. FOR IN VITRO USE ONLY.

Note that this data sheet is not lot-specific. Please consult the vial label and the certificate of analysis for information on specific lots.

Recombinant human RBM3

(Human Putative RNA-binding motif 3)

(Uniprot P98179)

Catalogue number: 12 110 002 Package size: 10 µg Catalogue number: 12 110 003 Package size: 200 µg

1. Protein characteristics

1.1 Molecular form: The RBM3 (Putative RNA-binding protein 3) is a full length, recombinant protein. The protein consists of 157 amino acids and a C-terminal His6-tag:

MSSEEGKLFVGGLNFNTDEQALEDHFSSFGPISEVVVVKDRETQRSRGFGFITFTNPEHASVAMRAMNGESLDG RQIRVDHAGKSARGTRGGGFGAHGRGRSYSRGGGDQGYGSGRYYDSRPGGYGYGYGRSRDYNGRNQGGYDR YSGGNYRDNYDNHHHHHH kDa

The calculated M of the His-tagged protein is 17.2 kDa. The protein is solubilized 70 r 55 in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2.

40

1.2 Purity: The recombinant RMB3 appears as a major band at about 17 kDa in SDS- 35

PAGE. It represents more than 90 % of total protein in the preparation. 25

1.3 Stability and storage: RBM3 is stable until the expiry date given on the label when stored at -70°C. The protein can be kept at -20°C for several weeks and on 15 ice for several days. Repeated freezing and thawing should be avoided. SDS-PAGE of 2. Applications 3 µg RBM3

The recombinant RBM3 can serve as standard in immunochemical assays, in Western Blot or other methods.

3. Introduction to RBM3

The cold shock protein RBM3 (RNA-binding motif 3) , encoding a 157-amino acid protein with a molecular weight of 17 kDa, can be stimulated in response to mild hypothermia (1, 2). Substantial evidence has shown that RBM3 acts as an important mediator of mild hypothermia in neuroprotection (3). In mouse models of degenerative diseases (4), RBM3 was found to mediate structural plasticity and protective effects of cooling against neuron loss. In vitro, RBM3 confers clear neuroprotective effects from apoptosis induced by various insults, such as serum and glucose deprivation, staurosporine, H2O2, nitric oxide (NO), retinoic acid (RA), and UV irradiation (5-9). As an RNA-binding protein, RBM3 may promote neural cell survival by accelerating ribosome assembly, affecting microRNA biosynthesis, stabilizing mRNA structure and increasing global de novo protein synthesis (10, 11). Another question has also been raised about the role of RBM3 in neurodegenerative diseases. Indeed, in 2018 was shown (12) that RBM3 confers neuroprotective effects in MPP+ (1-methyl-4-phenylpyridinium) -induced apoptosis in human neuroblastoma cell,

BioTeZ Berlin-Buch GmbH Robert-Rössle-Str. 10, 13125 Berlin, Germany Tel.: (0)30 / 9489-3322 Fax: (0)30 / 949-4509

Version March 2019

For research use only. Not for use in diagnostic procedures. FOR IN VITRO USE ONLY.

Note that this data sheet is not lot-specific. Please consult the vial label and the certificate of analysis for information on specific lots. thereby suggesting that RBM3-specific induction/overexpression might be used as a strategy for the treatment of neurodegenerative diseases. RBM3 was also found to elevate expression in different kinds of cancers and to be connected with clinic outcome, suggesting that RBM3 may be a potential favorable in clinical diagnoses. However, the expression level and clinical behavior of RBM3 in cancer is still conflicting, more regulation mechanisms of RBM3 in oncogenic or tumor suppressor are still in great need both in vitro and in vivo (13). Targeted as biomarker, there is a continuing need to assess the relationship of RBM3 expression level in different stages of various cancers to stratify patient for personalized prevention and therapy.

4. References

1. Derry, J. M., Kerns, J. A., and Francke, U. (1995). RBM3, a novel human gene in Xp11.23 with a putative RNA-binding domain. Hum.Mol. Genet. 4, 2307–2311. doi: 10.1093/hmg/4.12.2307 2. Danno, S., Nishiyama, H., Higashitsuji, H., Yokoi, H., Xue, J. H., Itoh, K., et al. (1997). Increased transcript level of RBM3, a member of the glycine-rich RNA-binding protein family, in human cells in response to cold stress. Biochem. Biophys. Res. Commun. 236, 804–807. doi: 10.1006/bbrc.1997. 7059 3. Chip, S., Zelmer, A., Ogunshola, O. O., Felderhoff-Mueser, U., Nitsch, C., Bührer, C., et al. (2011). The RNA-binding protein RBM3 is involved in hypothermia induced neuroprotection. Neurobiol. Dis. 43, 388–396. doi: 10.1016/j.nbd.2011.04.010 4. Peretti, D., Bastide, A., Radford, H., Verity, N., Molloy, C., Martin, M. G., et al. (2015). RBM3 mediates structural plasticity and protective effects of cooling in neurodegeneration. Nature 518, 236–239. doi: 10.1038/nature14142 5. Ferry, A. L., Vanderklish, P. W., and Dupont-Versteegden, E. E. (2011). Enhanced survival of skeletal muscle myoblasts in response to overexpression of cold shock protein RBM3. Am. J. Physiol. Cell Physiol. 301, 392–402. doi: 10.1152/ajpcell.00098.2011 6. Wellmann, S., Truss, M., Bruder, E., Tornillo, L., Eelmer, A., Seeger, K., et al. (2011). The RNA-binding protein RBM3 is required for cell proliferation and protects against serum deprivation-induced cell death. Pediatr. Res. 67, 35–41. doi: 10.1203/PDR.0b013e3181c13326 7. Ma, S. P., Ju, F., Zhang, Y. P., Shi, X., Zhuang, R. J., Xue, H., et al. (2017). Cold-inducible protein RBM3 protects neuroblastoma cells from retinoic acidinduced apoptosis via AMPK, p38 and JNK signaling. J. Funct. Foods 35,175–184. doi: 10.1016/j.jff.2017.05.045 8. Yang, H., Xia, Y., Lu, S. Q., Soong, T. W., and Feng, Z. W. (2008). Basic fibroblast growth factor-induced neuronal differentiation of mouse bone marrow stromal cells requires FGFR-1, MAPK/ERK, and transcription factor AP-1. J. Biol. Chem. 283, 5287–5295. doi: 10.1074/jbc.M706917200 9. Zhuang, R. J.,Ma, J., Shi, X., Ju, F.,Ma, S. P.,Wang, L., et al. (2017). Cold-inducible protein RBM3 protects UV irradiation-induced apoptosis in neuroblastoma cells by affecting p38 and JNK pathways and Bcl2 family . J. Mol. Neurosci. 63, 142–151. doi: 10.1007/s12031-017-0964-3 10. Zhou, T., Liang, Y., Jiang, L., Yu, T., Zeng, C., and Tao, E. (2017). Mild hypothermia protects against oxygen glucose deprivation/reoxygenation-induced apoptosis via the Wnt/b-catenin signaling pathway in hippocampal neurons. Biochem. Biophys. Res. Commun. 486, 1005–1013. doi: 10.1016/j.bbrc.2017.03.153 11. Zhu, X., Bührer, C., and Wellmann, S. (2016). Cold-inducible proteins CIRP and RBM3, a unique couple with activities far beyond the cold. Cell. Mol. Life Sci. 73, 3839–3859. doi: 10.1007/s00018-016-2253-7 12. Yang HJ, Shi X, Ju F, Hao BN, Ma SP, Wang L, Cheng BF, Wang M. (2018). Cold shock induced protein RBM3 but not mild hypothermia protects human SH-SY5Y neuroblastoma cells from MPP+-induced neurotoxicity. Front Neurosci.12:298. doi: 10.3389/fnins.2018.00298 13. Zhou, R. B., Lu, X. L., Zhang, C. Y., and Yin, D. C. (2017). RNA binding motif protein 3: a potential biomarker in cancer and therapeutic target in neuroprotection. Oncotarget 8, 22235–22250. doi: 10.18632/oncotarget.14755

BioTeZ Berlin-Buch GmbH Robert-Rössle-Str. 10, 13125 Berlin, Germany Tel.: (0)30 / 9489-3322 Fax: (0)30 / 949-4509

Version March 2019