| Purification and Quality Control | The soluble, His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified CAR is greater than 90% homogeneous based on SDS-PAGE analysis. |
| 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; 20 units are sufficient for in vitro transcription assay and 100 units are sufficient for protein-protein interaction assays. |
| Applications | CAR can be applied in in vitro transcription assays, DNA-protein and protein-protein interaction assays. For Research Use Only. |
| 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. |
| Synonyms | CAR; CAR1; MB67; MGC150433; MGC97144; MGC97209 and NR1I3. |
| Protein Sequence | MASREDELRN CVVCGDQATG YHFNALTCEG CKGFFRRTVS KSIGPTCPFA GSCEVSKTQR
RHCPACRLQK CLDAGMRKDM ILSAEALALR RAKQAQRRAQ QTPVQLSKEQ EELIRTLLGA
HTRHMGTMFE QFVQFRPPAH LFIHHQPLPT LAPVLPLVTH FADINTFMVL QVIKFTKDLP
VFRSLPIEDQ ISLLKGAAVE ICHIVLNTTF CLQTQNFLCG PLRYTIEDGA RVGFQVEFLE
LLFHFHGTLR KLQLQEPEYV LLAAMALFSP DRPGVTQRDE IDQLQEEMAL TLQSYIKGQQ
RRPRDRFLYA KLLGLLAELR SINEAYGYQI QHIQGLSAMM PLLQEICS |
| Background | The constitutive androstane receptor (CAR) was identified as a member of the orphan nuclear hormone receptor family in 1994 (1). Although constitutively active, it can be further activated by `phenobarbitol-like` compounds, the most potent being the synthetic compound TCPOBOP (2). Upon activation, CAR regulates the xenobiotic drug metabolizing enzymes, cytochrome P450s (2). There are several overlapping functions between the nuclear receptors PXR and CAR (3). Recently, CAR, as well as PXR, have been shown to place a role in bile acid clearance and cholestatic liver injury (4,5,6). |
| References | 1. Baes, M., et al. (1994) Mol. Cell Biol. 14: 1544-52
2. Wei, P., et al. (2000) Nature, 407; 920-23
3. Wei, P., et al. (2002) Pharmacogenomics J. 2: 117-26
4. Huang, W., et al. (2003) PNAS USA 100: 4156-61
5. Saini, S.P., et al. (2004) Mol. Pharmacol. 65: 292-300
6. Stedman, C.A., et al. (2005) PNAS USA 102: 2063-2068 |
| Image of SDS-PAGE /Western-blot |  |
