PEDF Human ELISA

Summary

PEDF is syntetized and released by human fetal retinal pigment epithelial cells (RPE) into the interphotoreceptor matrix and is localized to human chromosome 17p. It is a 50 kDa multifunctional glycoprotein belonging to the serpin protease inhibitor supergene (serpin) family, acting like substrates rather than inhibitors of serine proteases, being also described as serine peptidase inhibitor, clade F (alfa-2 antiplasmin, pigment epithelium derived factor), member 1. This gene encodes a 418 amino-acid protein with an asparagine glycosylation site at position 285–287 (Asn-Leu-Thr) and N-terminal signal peptide associated with secreted proteins. PEDF has an asymmetrical charge distribution, with a high density of basic residues concentrated on one side (positive) of the molecule and of acidic residues on the opposite side. Interactions of PEDF with three different types of molecules have been discovered: glycosaminoglycans of extracellular matrixes, collagens and receptors on the surface of neuronal cells. Negatively charged, acidic PEDF binds to collagen, lacks neurotrophic activity, and may confer antiangiogenic properties. PEDF has gliastatic, neuronotrophic, neuroprotective and antitumorigenic properties. PEDF acts in neuronal differentiation and survival in cells derived from retina and the central nervous system (CNS).Two functional epitopes have been identified on PEDF, a 34-mer peptide (residues 24–57) and a 44-mer peptide (residues 58–101). 44-mer peptide interacts with a a putative 80 kDa receptor (PEDFRN), identified on Y-79 cells (retinoblastoma cells), cerebellar and motor neurons, and in neural retina and replicates the neurotrophic function and the ability to block vascular leackage. The 34-mer peptide, possibly via a distinct receptor (PEDF-RA) identified on endothelial cells, induces apoptosis, blocks endothelial cell migration and corneal angiogenesis, but fails to induce Y-79 differentiation. Recently, PEDF was shown also to have potent anti-angiogenic activity as it specifically inhibited the migration of endothelial cells, an essential step in angiogenesis. Its activity equals or supersedes that of other anti-angiogenic factors, including angiostatin, endostatin and thrombospondin-1. In cell culture and in animal models, PEDF inhibited endothelial cell (EC) growth and migration and suppressed ischemia-induced neovascularization, whereas in porcine liver, the expression of PEDF has been associated with body muscularity and obesity. Analyses revealed that Human PEDF is correlated with BMI, CRP, diastolic blood pressure, insulin, Quicki. Individuals with metabolic syndrome (NCEP criterion) have significantly higher PEDF values than healthy subjects , suggesting that PEDF is and independent marker of MS with sufficient diagnostic efficacy.

Features

• It is intended for research use only.
• The total assay time is less than 4 hours.
• The kit measures PEDF in serum, plasma (EDTA, citrate, heparin).
• Assay format is 96 wells.
• Quality Controls are human serum based. No animal serum is used.
• Standard is recombinant protein based.
• Components of the kit are provided ready to use, concentrated or lyophilized.

Research topic

Energy metabolism and body weight regulation, Others

Assay format

Sandwich ELISA, Biotin-labelled antibody

Applications

Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum

Sample requirements

10 µl/well

Storage/Shipping

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

Calibration Curve

Calibration range

0.15 – 6 ng/ml

Limit of detection

0.045 ng/ml

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

CV = 3.57%

Inter-assay (Run-to-Run, n=5)

CV = 5.9%

Spiking Recovery

100.7%

Dilution Linearity

97.8%

Cross-Reactivity

References to this product

• Chen C, Tso AW, Law LS, Cheung BM, Ong KL, Wat NM, Janus ED, Xu A, Lam KS. Plasma level of pigment epithelium-derived factor is independently associated with the development of the metabolic syndrome in Chinese men: a 10-year prospective study. J Clin Endocrinol Metab. 2010 Nov;95 (11):5074-81
• Stejskal D, Karpisek M, Svestak M, Hejduk P, Sporova L, Kotolova H. Pigment epithelium-derived factor as a new marker of metabolic syndrome in Caucasian population. J Clin Lab Anal. 2010;24 (1):17-9
• Tschoner A, Sturm A, Ress C, Engl J, Kaser S, Laimer M, Laimer E, Klaus A, Tilg H, Patsch JR, Ebenbichler ChF. Effect of weight loss on serum pigment epithelium-derived factor levels. ECI. 2011 Jan;:6
• Yilmaz Y, Eren F, Ayyildiz T, Colak Y, Kurt R, Senates E, Tuncer I, Dolar E, Imeryuz N. Serum pigment epithelium-derived factor levels are increased in patients with biopsy-proven nonalcoholic fatty liver disease and independently associated with liver steatosis. Clin Chim Acta. 2011 Nov 20;412 (23-24):2296-9

References to summary

• Alberdi EM, Weldon JE, Becerra SP. Glycosaminoglycans in human retinoblastoma cells: heparan sulfate, a modulator of the pigment epithelium-derived factor-receptor interactions. BMC Biochem. 2003 Feb 19;4:1
• Amaral J, Becerra SP. Pigment Epithelium-Derived Factor and Angiogenesis. Retinal and Choroidal Angiogen. 19 January 2008;
• Aymerich MS, Alberdi EM, Martinez A, Becerra SP. Evidence for pigment epithelium-derived factor receptors in the neural retina. Invest Ophthalmol Vis Sci. 2001 Dec;42 (13):3287-93
• Becerra SP. Structure-Function Relationships of PEDF. National Eye Institute. December 2006;
• Becerra SP, Alberdi E, Enyadike C, Turgeon VL, Li L, D'Costa AP, Houenou LJ. Pigment epithelium-derived factor promotes the survival and differentiation of developing spinal motor neurons. J Comp Neurol. 1999 Sep 27;412 (3):506-14
• Becerra SP, Aymerich MS, Alberdi E. Binding of pigment epithelium-derived factor (PEDF) to retinoblastoma cells and cerebellar granule neurons. Evidence for a PEDF receptor. J Biol Chem. 1999 Oct 29;274 (44):31605-12
• Becerra SP, Notari L, Meyer C. Mapping the type I collagen-binding site on pigment epithelium-derived factor. Implications for its antiangiogenic activity. J Biol Chem. 2002 Nov 22;277 (47):45400-7
• Becerra SP, Palmer I, Kumar A, Steele F, Shiloach J, Notario V, Chader GJ. Overexpression of fetal human pigment epithelium-derived factor in Escherichia coli. A functionally active neurotrophic factor.
• Becerra SP, Sagasti A, Spinella P, Notario V. Pigment epithelium-derived factor behaves like a noninhibitory serpin. Neurotrophic activity does not require the serpin reactive loop.
• Cai J, Jiang WG, Grant MB, Boulton M. Pigment epithelium-derived factor inhibits angiogenesis via regulated intracellular proteolysis of vascular endothelial growth factor receptor 1. J Biol Chem. 2006 Feb 10;281 (6):3604-13
• Cleeman JI, Daniels SR, Donato KA, Eckel RH, Franklin BA, Gordon DJ, Krauss RM, Savage PJ, Smith SC Jr, Spertus JA, Fernando Costa, Grundy SM. Diagnosis and management of the metabolic syndrome: an American Heart Association/National Heart, Lung, and Blood Institute scientific statement: Executive Summary. Crit Pathw Cardiol. 2005 Dec;4 (4):198-203
• Ek ET, Dass CR, Choong PF. Pigment epithelium-derived factor: a multimodal tumor inhibitor. Mol Cancer Ther. 2006 Jul;5 (7):1641-6
• Filleur S, Volz K, Nelius T, Mirochnik Y, Huang H, Zaichuk TA, Aymerich MS, Becerra SP, Yap R, Veliceasa D, Shroff EH, Volpert OV. Two functional epitopes of pigment epithelial-derived factor block angiogenesis and induce differentiation in prostate cancer. Cancer Res. 2005 Jun 15;65 (12):5144-52
• Gettins PG, Simonovic M, Volz K. Pigment epithelium-derived factor (PEDF), a serpin with potent anti-angiogenic and neurite outgrowth-promoting properties.
• Greenberg J, Goliath R, Tombran-Tink J, Rodriquez IR, Ramesar R, Chader G. The gene for PEDF, a retinal growth factor is a prime candidate for retinitis pigmentosa and is tightly linked to the RP13 locus on chromosome 17p13.3. Mol Vis. 1996 Jun 19;2:5
• Imaizumi T, Nakamura K, Matsui T, Takenaka K, Jinnouchi Y, Hino A, Furuki K, Enomoto M, Yashiro T, Abe A, Adachi H, Yamagishi S. Elevated serum levels of pigment epithelium-derived factor in the metabolic syndrome. J Clin Endocrinol Metab. 2006 Jun;91 (6):2447-50
• Liu HY, Gu Q, Fan Y, Huang JN, Zhu Q, Xu X, Zhang X, Sun XD, Wang FH. Role of pigment epithelium-derived factor on proliferation and migration of choroidal capillary endothelium induced by vascular endothelial growth factor in vitro. Chin Med J (Engl). 2007 Sep 5;120 (17):1534-8
• Lyons TJ, Harper CA, Dragicevic G, Mori TA, Croft KD, Januszewski AS, O'Neal DN, Nelson CL, Chung JS, Rowley KG, Karschimkus CS, Zhang SX, Jenkins AJ, Best JD, Ma JX. Increased serum pigment epithelium-derived factor is associated with microvascular complications, vascular stiffness and inflammation in Type 1 diabetes. Diabet Med. 2007 Dec;24 (12):1345-51
• Ma JX, Lau K, Cao W, Knapp RR, Mott R, Zhang SX, Wang JJ. Salutary effect of pigment epithelium-derived factor in diabetic nephropathy: evidence for antifibrogenic activities. Diabetes. 2006 Jun;55 (6):1678-85
• Matsunaga N, Chikaraishi Y, Izuta H, Ogata N, Shimazawa M, Matsumura M, Hara H. Role of soluble vascular endothelial growth factor receptor-1 in the vitreous in proliferative diabetic retinopathy. Ophthalmology. 2008 Nov;115 (11):1916-22
• Motomiya Y, Yamagishi S, Adachi H, Abe A. Increased serum concentrations of pigment epithelium-derived factor in patients with end-stage renal disease. Clin Chem. 2006 Oct;52 (10):1970-1
• Petersen SV, Nielsen SB, Valnickova Z, Enghild JJ, Otzen DE. Heparin binding induces a conformational change in pigment epithelium-derived factor. J Biol Chem. 2007 Mar 2;282 (9):6661-7
• Petersen SV, Valnickova Z, Enghild JJ. Pigment-epithelium-derived factor (PEDF) occurs at a physiologically relevant concentration in human blood: purification and characterization.
• Pfeffer BA, Wong P, Montuenga LM, Wu YQ, Fariss RN, Becerra SP. Pigment epithelium-derived factor in the monkey retinal pigment epithelium and interphotoreceptor matrix: apical secretion and distribution. Exp Eye Res. 2004 Feb;78 (2):223-34
• Shao H, Schvartz I, Shaltiel S. Secretion of pigment epithelium-derived factor. Mutagenic study. Eur J Biochem. 2003 Mar;270 (5):822-31
• Simonovic M, Gettins PG, Volz K. Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor.
• Stratikos E, Alberdi E, Gettins PG, Becerra SP. Recombinant human pigment epithelium-derived factor (PEDF): characterization of PEDF overexpressed and secreted by eukaryotic cells.
• Xu X, Zhang SS, Barnstable CJ, Tombran-Tink J. Molecular phylogeny of the antiangiogenic and neurotrophic serpin, pigment epithelium derived factor in vertebrates. BMC Genomics. 2006;7:248

By research topic:

• Energy metabolism and body weight regulation
• Others

By molecule:

• Pigment Epithelium-Derived Factor

Registration form