ELISA Kit for Pigment Epithelium Derived Factor (PEDF) Rattus norvegicus (Rat) Sandwich ELISA

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

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  • ELISA Kit for Pigment Epithelium Derived Factor (PEDF) Packages (Simulation)
  • ELISA Kit for Pigment Epithelium Derived Factor (PEDF) Packages (Simulation)
  • ELISA Kit for Pigment Epithelium Derived Factor (PEDF) Results demonstration
  • SEB972Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-93 83
EDTA plasma(n=5) 87-101 97
heparin plasma(n=5) 88-99 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pigment Epithelium Derived Factor (PEDF) 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) 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) 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) 84-99% 86-93% 96-105% 93-104%
EDTA plasma(n=5) 91-103% 89-101% 82-93% 95-105%
heparin plasma(n=5) 88-101% 87-96% 87-94% 78-102%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Pigment Epithelium Derived Factor (PEDF)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Pigment Epithelium Derived Factor (PEDF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Pigment Epithelium Derived Factor (PEDF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Pigment Epithelium Derived Factor (PEDF), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Pigment Epithelium Derived Factor (PEDF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • A Therapeutic Strategy for Choroidal Neovascularization Based on Recruitment of Mesenchymal Stem Cells to the Sites of LesionsNature: 2010144
  • Urinary Pigment Epithelium-Derived Factor as a Marker of Diabetic NephropathyKarger: 314326
  • Increased expression of pigment epithelium-derived factor in aged mesenchymal stem cells impairs their therapeutic efficacy for attenuating myocardial infarction injuryOxfordJournals: source
  • The change of serum level of pigment epithelium-derived factor in coronary heart diseaseBmj: Source
  • Metformin Inhibits Expression and Secretion of PEDF in Adipocyte and Hepatocyte via Promoting AMPK PhosphorylationPubmed: 24288442
  • The association study of plasma levels of pigment epithelium-derived factor with acute coronary syndrome in the chinese han population.Pubmed: 24192856
  • Rosiglitazone Inhibits Expression and Secretion of PEDF in Adipose Tissue and Liver of Male SD Rats Via a PPAR-γ Independent MechanismEndocrine: Source
  • Pigment epithelium-derived factor regulates microvascular permeability through adipose triglyceride lipase in sepsisPubMed: 25700221
  • Pigment Epithelium-Derived Factor (PEDF) Protects Osteoblastic Cell Line from Glucocorticoid-Induced Apoptosis via PEDF-RPubmed:27187377
  • The Ingenious Interactions Between Macrophages and Functionally Plastic Retinal Pigment Epithelium Cellsarticleid:2582822
  • VEGF-B promotes recovery of corneal innervations and trophic functions in diabetic micepubmed:28091556
  • 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
  • PEDF Reduces the Severity of Herpetic Simplex Keratitis in MicePubmed:30025136
  • 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
  • Lactation-related changes in tissue expression of PEDF in dairy cowsPubmed:29758402
  • Correlation Between Pigment Epithelium-Derived Factor (PEDF) level and Degree of Coronary Angiography and Severity of Coronary Artery Disease in a …Pubmed:29574467
  • Role of pigment epithelium-derived factor (PEDF) on arsenic-induced neuronal apoptosisDoi: 10.1016/j.chemosphere.2018.10.100
  • Role of Pigment Epithelium-Derived Factor in Arsenic-Induced Vascular Endothelial Dysfunction in a Rat ModelPubmed: 30392020
  • Increased levels of serum pigment epithelium-derived factor aggravate proteinuria via induction of podocyte actin rearrangementPubmed: 30536192
  • A novel xeno-free culture system for human retinal pigment epithelium cellsPubmed: 31024807
  • Pigment Epithelial‐Derived Factor Deficiency Accelerates Atherosclerosis Development via Promoting Endothelial Fatty Acid Uptake in Mice With HyperlipidemiaPubmed: 31711388
  • Pigment epithelium-derived factor (PEDF) ameliorates arsenic-induced vascular endothelial dysfunction in rats and toxicity in endothelial EA. hy926 cellsPubmed: 32315827
  • The Expression Levels and Cellular Localization of Pigment Epithelium Derived Factor (PEDF) in Mouse Testis: Its Possible Involvement in the Differentiation of Spermatogonial Cells33498962
  • 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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