Wide-range ELISA Kit for Endostatin (ES)

Specificity

This assay has high sensitivity and excellent specificity for detection of Wide-range Endostatin (ES).
No significant cross-reactivity or interference between Wide-range Endostatin (ES) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Wide-range Endostatin (ES) and the recovery rates were calculated by comparing the measured value to the expected amount of Wide-range Endostatin (ES) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-102 88
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 80-97 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wide-range Endostatin (ES) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Wide-range Endostatin (ES) 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 Wide-range Endostatin (ES) 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) 96-105% 94-101% 97-104% 81-98%
EDTA plasma(n=5) 82-97% 89-98% 97-105% 80-98%
heparin plasma(n=5) 88-105% 86-94% 96-105% 95-105%

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.

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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
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