Certificate of Analysis and Data Sheet
Source:
E.coli
Catalog No.
CYT-437
Introduction:
Clusterin also names Apoliprotein J is a 75-80 kD disulfide-linked heterodimeric protein containing about 30% of N-linked carbohydrate rich in sialic acid but truncated forms targeted to the nucleus have also been identified.
The precursor polypeptide chain is cleaved proteolytically to remove the 22-mer secretory signal peptide and subsequently between residues 227/228 to generate the alpha and beta chains. These are assembled in anti-parallel to give a heterodimeric molecule in which the cysteine-rich centers are linked by five disulfide bridges and are flanked by two predicted coiled-coil alpha-helices and three predicted amphipathic a-helices.
Across a broad range of species clusterin shows a high degree of sequence homology ranging from 70% to 80%. It is nearly ubiqitously expressed in most mammalian tissues and can be found in plasma, milk, urine, cerebrospinal fluid and semen.
It is able to bind and form complexes with numerous partners such as immunoglobulins, lipids, heparin, bacteria, complement components, paraoxonase, beta amyloid, leptin and others. Clusterin has been ascribed a plethora of functions such as phagocyte recruitment, aggregation induction, complement attack prevention, apoptosis inhibition, membrane remodelling, lipid transport, hormone transport and/or scavenging, matrix metalloproteinase inhibition.
A genuine function of clusterin has not been defined. One tempting hypothesis says that clusterin is an extracellular chaperone protecting cells from stress induced insults caused by degraded and misfolded protein precipitates.
Clusterin is up- or downregulated on the mRNA or protein level in many pathological and clinically relevant situations including cancer, organ regeneration, infection, Alzheimer disease, retinitis
pigmentosa, myocardial infarction, renal tubular damage, autoimmunityand others.
Description:
Recombinant Rat Clusterin was constructed as a recombinant protein with N-terminal fusion of T7-Tag (16AA) and C-terminal fusion of His-Tag (9AA). The Rat Clusterin His-Tagged Fusion Protein, produced in E.coli, is 26.5 kDa protein containing 215 amino acid residues of the Rat Clusterin and 25 additional amino acid residues ?His-Tag, T7-Tag (bold).
MASMTGGQQM GRDPNSSSPF YFWMNGDRID SLLESDRQQS QVLDAMQDSF TRASGIIDTL FQDRFFTHEPQDIHHFSPMG FPHKRPHLLY PKSRLVRSLM PLSHYGPLSF HNMFQPFFDM IHQAQQAMDV QLHSPALQFPDVDFLKEGED DRTVCKEIRH NSTGCLKMKG QCEKCQEILS VDCSTNNPAQ ANLRQELNDS LQVAERLTQQYNELLHSLQS KMLNTSSLLE Q
ALEHHHHHH
Specificity:
The amino acid sequence of the recombinant Rat Clusterin is 100% homologous to the amino acid sequence 146-360 of the Rat Clusterin precursor.
Purification Method:
Ni-NTA chromatography
Purity:
>95% (SDS PAGE analyzed).
Formulation:
Sterile filtered and lyophilized from 0.5 mg/ml in 0.01M Tris pH 7.2.
Reconstitution:
Add 0.2 ml of deionized H
2O and let the lyophilized pellet dissolve completely.
Storage:
Store lyophilized protein at -20蚓. Aliquot the product after reconstitution to
avoid repeated freezing/thawing cycles. Reconstituted protein can be stored at
4蚓 for a limited period of time; it does not show any change after two weeks at
4蚓.
Please avoid freeze-thaw cycles.
Stability/Shelf Life:
The lyophilized protein remains stable until the expiry date when stored at -20蚓.
Applications:
Western blotting
References:
?Trougakos IP, Gonos ES. Functional analysis of clusterin/apolipoprotein J in cellular death induced by severe genotoxic stress.Ann N Y Acad Sci. 2004 Jun;1019:206-10.
?Carver JA, Rekas A, Thorn DC, Wilson MR. Small heat-shock proteins and clusterin: intra- and extracellular molecular chaperones with a common mechanism of action and function? IUBMB Life. 2003 Dec; 55(12): 661-8. Review.
?Min BH, Kim BM, Lee SH, Kang SW, Bendayan M, Park IS. Clusterin expression in the early process of pancreas regeneration in the pancreatectomized rat. J Histochem Cytochem. 2003 Oct; 51(10): 1355-65.
?Park JH, Park JS, Ju SK, Lee KB, Park YK, Kang MH, Na SY, You KH. Clusterin mRNA expression in apoptotic and activated rat thymocytes. Cell Res. 2003 Feb; 13(1): 49-58.
Usage:
This material is offered by ProSpec-TechnoGene for research,
laboratory or further evaluation purposes.
Latest Publications:
1. Clusterin is a secreted marker for a hypoxia-inducible factor-independent function of the von hippel-lindau tumor suppressor protein.
Am J Pathol 2006 Feb;168(2):574-84
2. Value of clusterin immunoreactivity as a predictive factor in muscle-invasive urothelial bladder carcinoma.
Urology 2006 Jan;67(1):105-9
3. Clusterin induces differentiation of pancreatic duct cells into insulin-secreting cells.
Diabetologia 2006 Jan 13;:1-10
4. Knock-down of the Cytoprotective Gene, Clusterin, to Enhance Hormone and Chemosensitivity in Prostate and Other Cancers.
Ann N Y Acad Sci 2005 Nov;1058:1-15
5. Knockdown of the cytoprotective chaperone, clusterin, chemosensitizes human breast cancer cells both in vitro and in vivo.
Mol Cancer Ther 2005 Dec;4(12):1837-49
6. Up-regulation of the clusterin gene after proteotoxic stress. Implication of HSF1/HSF2 heterocomplexes.
Biochem J 2005 Dec 9;