| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified HMG1 is greater than 95% homogeneous based on SDS-PAGE analysis. Endotoxin is removed during chromatography purification process. Low endotoxin levels, <0.25 EU/ml by LAL (Limulus Amebocyte Lysate Testing) using validation requirements established by the FDA. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 1 unit is sufficient for a gel mobility shift assay in a 20 µl reaction; 100 units are sufficient for protein-protein interaction assays. | | Applications | HMG1 can be applied in in vitro transcription assays, DNA-protein and protein-protein interactions assays. | | Formulation and Storage | The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles. | | Synonym | HMG1; HMG3; SBP-1. | | Protein Sequence | GKGDPKKPRG KMSSYAFFVQ TCREEHKKKH PDASVNFSEF SKKCSERWKT MSAKEKGKFE
DMAKADKARY EREMKTYIPP KGETKKKFKD PNAPKRPPSA FFLFCSEYRP KIKGEHPGLS
IGDVAKKLGE MWNNTAADDK QPYEKKAAKL KEKYEKDIAA YRAKGKPDAA KKGVVKAEKS
KKKKEEEEDE EDEEDEEEEE DEEDEDEEED DDDE | | Background | High mobility group 1 (HMG1) is a 26 kDa highly conserved non-sequence-specific DNA-binding nuclear protein (1). Mammalian HMG1 has two homologous DNA-binding domains: HMG boxes, A and B (each of 80–90 amino-acid residues), linked by a short basic region to an acidic C-terminal domain containing 30 consecutive Asp and Glu residues (2). HMG1 has been implicated in a number of fundamental biological processes including transcription, replication and recombination, in which it plays a role in manipulating DNA structure by bending, looping, compacting or unwinding, or by direct contacts with distinct cellular proteins (3-5). HMG-1 can act as a repressor, by interacting with TBP to block pre-initiation complex formation (6, 7) or as an activator, by facilitating the binding of various transcription factors to their cognate DNA sequences (8, 9). Most recently, it was discovered that HMG-1 is a late mediator of delayed endotoxin lethality by activating downstream cytokine release (10).
Recombinant His tagged HMG1 is isolated from an E. coli strain that carries the coding sequence of the human HMG1 under the control of a T7 promoter. | | References | 1. Bianchi, M.E., et al., (1989) Science 243, 1056-1059
2. Bustin, M., et al., (1990) Biochim. Biophys. Acta 1049, 231-243
3. Zappavigna, V., et al., (1996) EMBO J. 15, 4981-4991
4. Ge, H., et al., (1994) J. Biol. Chem. 269, 17136-17140
5. Zlatanova, Y. et al., (1998) FASEB J. 12, 421-431
6. Stelzer, G., et al., (1994) Mol. Cell. Biol. 14, 4712-4721
7. Lu, W., et al., (2000) J. Biol. Chem. Nov 10;275(45):35006-12
8. Jayaraman, L., et al., (1998) Genes Dev. 12, 462-472
9. Onate, S., et al., (1994) Mol. Cell. Biol. 14, 3376-3391
10. Yang, H., et al., (2001) Shock 15, 247-253 | | Image of SDS-PAGE /Western-blot |  |
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