Multiplex Assay Kit for Endostatin (ES) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 96
EDTA plasma(n=5) 81-105 98
heparin plasma(n=5) 87-103 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Endostatin (ES) ,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 Endostatin (ES) ,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 Endostatin (ES) ,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) 80-103% 78-94% 81-89% 91-102%
EDTA plasma(n=5) 87-94% 80-97% 93-102% 96-104%
heparin plasma(n=5) 93-105% 86-104% 78-90% 94-101%

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#:ES) 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
Cancer Immunology Endostatin gene therapy enhances the efficacy of IL-2 in suppressing metastatic renal cell carcinoma in mice SpringerLink: u38777121t6660l5
Journal of Fluoresence Erythrocyte Protoporphyrin Fluorescence as a Biomarker for Monitoring Antiangiogenic Cancer Therapy SpringerLink: 4v306130302257w0
The Journal of Gene Medicine Endostatin- and interleukin-2-expressing retroviral bicistronic vector for gene therapy of metastatic renal cell carcinoma Wiley: source
Biomedicine & Pharmacotherapy Endostatin neoadjuvant gene therapy extends survival in an orthotopic metastatic mouse model of renal cell carcinoma ScienceDirect: S075333221100151X
Science Alert Antiangiogenic Activities of Cinnamon, Black and Green Tea Extracts on Experimentally Induced Breast Cancer in Rats Scialert: Source
The Journal of Immunology Inflammation Induced by MMP-9 Enhances Tumor Regression of Experimental Breast Cancer PubMed: PMC3619527
Prostaglandins &amp; Other Lipid Mediators Promotion of adipogenesis by an EP2 receptor agonist via stimulation of angiogenesis in pulmonary emphysema Pubmed:24911647
American Journal of Physiology - Heart and Circulatory Physiology Endostatin and kidney fibrosis in aging: a case for antagonistic pleiotropy? Source
shock Angiogenesis inhibitor endostatin protects mice with sepsis from multiple organ dysfunction syndrome PubMed: 26125086
Clin Rev Allergy Immunol Endothelial Dysfunction and Nailfold Videocapillaroscopy Pattern as Predictors of Digital Ulcers in Systemic Sclerosis: a Cohort Study and Review of the Literature PubMed: 26142066
Angiologia e Cirurgia Vascular Peripheral vasculopathy in Raynaud phenomenon: Vascular disease biomarkers science:S1646706X16300052
Clinical Rheumatology Impaired angiogenesis as a feature of digital ulcers in systemic sclerosis Pubmed:26920752
Blood biomarker for early blood brain barrier disruption in ischemic stroke patent:US20170199205A1
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