Multiplex Assay Kit for Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay)

SERPINF1; EPC1; PIG35; SDF3; Serpin F1; Cell proliferation-inducing gene 35; Serpin Peptidase Inhibitor,Clade F Member 1(Alpha-2 Antiplasmin); Stromal Cell Derived Factor 3

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-98 95
EDTA plasma(n=5) 80-103 92
heparin plasma(n=5) 97-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Pigment Epithelium Derived Factor (PEDF) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 96-103% 82-101% 93-105% 89-96%
EDTA plasma(n=5) 78-91% 88-99% 82-99% 84-94%
heparin plasma(n=5) 94-102% 82-101% 94-102% 80-97%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:PEDF) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Molecular Therapy A Therapeutic Strategy for Choroidal Neovascularization Based on Recruitment of Mesenchymal Stem Cells to the Sites of Lesions Nature: 2010144
American Journal of Nephrology Urinary Pigment Epithelium-Derived Factor as a Marker of Diabetic Nephropathy Karger: 314326
34 Increased expression of pigment epithelium-derived factor in aged mesenchymal stem cells impairs their therapeutic efficacy for attenuating myocardial infarction injury OxfordJournals: source
Heart and Education in Heart The change of serum level of pigment epithelium-derived factor in coronary heart disease Bmj: Source
Mediators of Inflammation Metformin Inhibits Expression and Secretion of PEDF in Adipocyte and Hepatocyte via Promoting AMPK Phosphorylation Pubmed: 24288442
Cardiology. The association study of plasma levels of pigment epithelium-derived factor with acute coronary syndrome in the chinese han population. Pubmed: 24192856
Endocrinology Rosiglitazone Inhibits Expression and Secretion of PEDF in Adipose Tissue and Liver of Male SD Rats Via a PPAR-γ Independent Mechanism Endocrine: Source
Clinical Science Pigment epithelium-derived factor regulates microvascular permeability through adipose triglyceride lipase in sepsis PubMed: 25700221
Molecular Sciences Pigment Epithelium-Derived Factor (PEDF) Protects Osteoblastic Cell Line from Glucocorticoid-Induced Apoptosis via PEDF-R Pubmed:27187377
Investigative ophthalmoloey & visual science The Ingenious Interactions Between Macrophages and Functionally Plastic Retinal Pigment Epithelium Cells articleid:2582822
Scientific reports VEGF-B promotes recovery of corneal innervations and trophic functions in diabetic mice pubmed:28091556
国际眼科杂志 色素上皮衍生因子修饰的人脐带间充质干细胞构建方法 201707007
Investigative Ophthalmology & Visual Science PEDF Reduces the Severity of Herpetic Simplex Keratitis in Mice Pubmed:30025136
International Eye Science 高海拔地区新生血管性青光眼不同治疗方式的疗效及对房水中PEDF, VEGF 水平的影响 2018/2/201802020.pdf
Domestic Animal Endocrinology Lactation-related changes in tissue expression of PEDF in dairy cows Pubmed:29758402
Medical Science Monitor Correlation Between Pigment Epithelium-Derived Factor (PEDF) level and Degree of Coronary Angiography and Severity of Coronary Artery Disease in a … Pubmed:29574467
Chemosphere Role of pigment epithelium-derived factor (PEDF) on arsenic-induced neuronal apoptosis Doi: 10.1016/j.chemosphere.2018.10.100
Biological Trace Element Research Role of Pigment Epithelium-Derived Factor in Arsenic-Induced Vascular Endothelial Dysfunction in a Rat Model Pubmed: 30392020
International Urology and Nephrology Increased levels of serum pigment epithelium-derived factor aggravate proteinuria via induction of podocyte actin rearrangement Pubmed: 30536192
International Journal of Ophthalmology A novel xeno-free culture system for human retinal pigment epithelium cells Pubmed: 31024807
Journal of the American Heart Association Pigment Epithelial‐Derived Factor Deficiency Accelerates Atherosclerosis Development via Promoting Endothelial Fatty Acid Uptake in Mice With Hyperlipidemia Pubmed: 31711388
environmental research Pigment epithelium-derived factor (PEDF) ameliorates arsenic-induced vascular endothelial dysfunction in rats and toxicity in endothelial EA. hy926 cells Pubmed: 32315827
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES The Expression Levels and Cellular Localization of Pigment Epithelium Derived Factor (PEDF) in Mouse Testis: Its Possible Involvement in the Differentiation of Spermatogonial Cells 33498962
Hum Cell Long non-coding RNA Rian promotes the expression of tight junction proteins in endothelial cells by regulating perivascular-resident macrophage-like melanocytes ¡­ 33768511
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