Clusterin Human ELISA

Research topic

Others, Renal disease

Features

• It is intended for research use only.
• The total assay time is less than 3.5 hours.
• The kit measures Clusterin in serum, plasma, cerebrospinal fluid (CSF), urine, cell culture, and cell lysate.
• Components of the kit are provided ready to use, concentrated or lyophilized.
• Standard is recombinant protein based.
• Quality Controls are human serum based. No animal sera are used.

Storage/Shipping

Store the kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box).

Summary

Clusterin (Apolippoprotein J; SP-40,40; TRPM-2; SGP-2; pADHC-9; CLJ; T64; GP III; XIP8) is a highly conserved disulfide-linked secreted heterodimeric glycoprotein of 75–80 kDa but truncated forms targeted to the nucleus have also been identified. The protein is constitutively secreted by a number of cell types including epithelial and neuronal cells and is a major protein in physiological fluids including plasma, milk, urine, cerebrospinal fluid and semen. Due to its wide breath of tissue distribution many diverse physiological functions have been attributed to Clusterin including sperm maturation, membrane recycling, lipid transportation, tissue remodelling, complement inhibition and cell-cell or cell-substratum interactions. Moreover, it was proposed that Clusterin functions as an extracellular chaperon that stabilizes stressed proteins in a folding-competent state, and that the protein is involved in programmed cell death. Another defining prominent function of Clusterin is its induction in many severe physiological disturbances states including kidney degenerative diseases, prostate and vesicle carcinogenesis, ovarian cancer and several neurodegenerative conditions (Alzheimer’s di­sease). Recent study demonstrates, that serum Clusterin level increases significantly in diabetic type II patients and in patients with developing coronary heart disease, or myocardial infarction. These data raise the possibility that elevated Clusterin levels in serum may represent a strong indication of vascular damage. In patients with systemic lupus erythematosus (SLE), reduced serum Clusterin levels correlated inversely with disease activity. Lowered Clusterin levels could be involved in the pathogeneses of SLE on account of decreased protective effects. Another interesting observation obtained in rat model suggests that measurement of urinary Clusterin levels may be a useful clinical marker for the severity of renal tubular damage. Furthermore, urinary Clusterin may also help to differentiate between tubular and glomerular forms of proteinuria.

Areas of investigation: Coronary heart diseases, Myocardial infarction, Neurodegenerative diseases, Kidney degenerative disease, Renal tubular damage

Assay format

Sandwich ELISA, Biotin-labelled antibody

Sample requirements

5 µl/well

Applications

Cell lysate, Cerebrospinal fluid, Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum, Urine

Calibration Curve

Calibration range

5 to 160 ng/ml

Limit of detection

Limit of detection (LOD) (defined as concentration of analyte giving absorbance higher than mean absorbance of blank* plus three standard deviations of the absorbance of blank: Ablank + 3×SDblank) is calculated from the real human Clusterin values in wells and is: 0.5 ng/ml.

Limit of quantification

Limit of assay Results exceeding human Clusterin level of 160 ng/ml should be repeated with more diluted samples. Dilution factor needs to be taken into consideration in calculating the Clusterin concentration. Results from assaying of urine samples exceeding human Clusterin level of 40 ng/ml should be repeated with more diluted urine samples for correct values. Dilution factor needs to be taken into consideration in calculating the human Clusterin concentration.

Intra-assay (Within-Run, n=8)

CV = 6.2 %

Inter-assay (Run-to-Run, n=8)

CV = 7.8 %

Spiking Recovery

102.8 %

Dilution Linearity

98.3 %

Cross-Reactivity

Human, Monkey

Find out more on biovendor.com

• Clusterin in BioVendor Reviews

References to this product

• Stejskal D, Fiala RR . Evaluation of serum and urine clusterin as a potential tumor marker for urinary bladder cancer. Neoplasma . 53(4):343-6 (2006)
• Stejskal D, Lacnak B, Karpisek M, Kaminek M . Our experiences with measurement of new potential biomarkers in the diagnosis of latent forms of myocardial ischemia. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub . Dec;150(2):239-44 (2006)

References to summary

• Choi-Miura NH, Oda T: Relationship between multifunctional protein Clusterin and Alzheimer disease. Neurobiol. Aging 1996; 17(5): 717–722
• Newkirk MM, Apostolakos P, Neville C and Fortin PR: Systemic lupus erythematosus, a disease associated with low levels of Clusterin/ApoJ, and anti-inflammatory protein. J. Rheumatol.1999; 3:597–603
• Morrissey C, Lakins J, Moquin A, Hussain M, Tenniswood M: An antigen capture assay for the measurement of serum Clusterin concentrations. J. Biochen. Biophys. Methods 2001; 48:13–21
• Trougakos IP, Poulakou M, Stathatos M, Chalikia A, Melidonis A, Gonos ES: Serum levels of the senescence biomarker Clusterin/apo­lipoprotein J increase significantly in diabetes type II and during development of coronary heart disease or at myocardial infarction. Ex. Gerontology 2002; 37: 1175–1187
• Jones SE, Jomary C: Molecules in focus Clusterin. The International J. of Bioch. & Cell Biol. 2002; 34:427–431
• Hidaka S, Kränzlin B, Gretz N, Witzgall R: Urinary Clusterin levels in the rat correlate with the severity of tubular damage and may help to differentiate between glomerular and tubular injuries. Cell Tissue Res. 2002; 310:289–296
• Chen X, Halberg RB, Ehrhardt WM, Torrealba J and Dove WF: Clusterin as a biomarker in murine and human intestinal neoplasia. PNAS 2003; 100:9530–9535
• Zhang LY, Ying WT, Mao YS, He HZ, Liu Y, Wang HX, Liu F, Wang K, Zhang DC, Wang Y, Wu M, Qian XH and Zhao XH: Loss of Clusterin both in serum and tissue correlates with the tumorogenesis of esophageal squamous cell carcinoma via proteomics approaches. World J Gastroenterol 2003; 9:650–654
• Wang L, Erling P, Bengtsson AA, Truedsson L, Sturfelt G, Erlinge D: Transcriptional down-regulation of the platelet ADP receptory P2Y12 and Clusterin in patients with systemic lupus erythematosus. J. of Thromb. And Haemost. 2004; 2:1436–1442
• Patel NV, Wei M, Wong A, Finch CE, Morgan TE: Progressive changes in regulation of apolipoproteins E and J in glial cultures during postnatal development and aging. Neuroscience Letters 2004; 371:199–204
• Kim BM, Kim SY, Lee S, Shin YJ, Min BH, Bendayan M, Park IS: Clusterin induces differentiation of pancreatic duct calls into insulin-secreting cells. Diabetologia 2006; 49:311–320
• Kruger S, Mahnken A, Kausch I, Feller AC: Value of Clusterin immunoreactivity as a predictive factor in muscle-invasive urothelial bladder carcinoma. Urology 2006; 67:105–109
• Stejskal D, Fiala R: Evaluation of serum and urine Clusterin as a potential tumor marker for urinary bladder cancer. Neoplasma 2006; 53:343–34