ELISA Kit for Angiopoietin Like Protein 2 (ANGPTL2) Mus musculus (Mouse) Sandwich ELISA

HARP; ARP2; Angiopoietin-Related Protein 2

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  • ELISA Kit for Angiopoietin Like Protein 2 (ANGPTL2) Packages (Simulation)
  • ELISA Kit for Angiopoietin Like Protein 2 (ANGPTL2) Packages (Simulation)
  • ELISA Kit for Angiopoietin Like Protein 2 (ANGPTL2) Results demonstration
  • SEB919Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Angiopoietin Like Protein 2 (ANGPTL2) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiopoietin Like Protein 2 (ANGPTL2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-103 94
EDTA plasma(n=5) 90-98 94
heparin plasma(n=5) 91-103 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiopoietin Like Protein 2 (ANGPTL2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiopoietin Like Protein 2 (ANGPTL2) 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 Angiopoietin Like Protein 2 (ANGPTL2) 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) 94-102% 92-101% 84-98% 93-102%
EDTA plasma(n=5) 93-105% 93-102% 83-103% 90-104%
heparin plasma(n=5) 84-102% 83-90% 79-103% 81-89%

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 Angiopoietin Like Protein 2 (ANGPTL2)

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 Angiopoietin Like Protein 2 (ANGPTL2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Angiopoietin Like Protein 2 (ANGPTL2). 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 Angiopoietin Like Protein 2 (ANGPTL2), 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 Angiopoietin Like Protein 2 (ANGPTL2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Tumor necrosis factor-α increases angiopoietin-like protein 2 gene expression by activating Foxo1 in 3T3-L1 adipocytesPubMed: 21501655
  • Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditionsPubmed:24626957
  • Lack of Angiopoietin‐Like‐2 Expression Limits the Metabolic Stress Induced by a High‐Fat Diet and Maintains Endothelial Function in MicePubmed:25128474
  • Elevated Serum Angiopoietin-like Protein 2 Correlates with the Metastatic Properties of Colorectal Cancer: A Serum Biomarker for Early Diagnosis and RecurrencePubmed:25294915
  • Serum angiopoietin-like protein 2 as a potential biomarker for diagnosis, early recurrence and prognosis in gastric cancer patientsPubMed: 26420253
  • ANGPTL2 is associated with an increased risk of cardiovascular events and death in diabetic patients.pubmed:27491833
  • c-kit Positive Cardiac Outgrowth Cells Demonstrate Better Ability for Cardiac Recovery Against Ischemic Myopathy.pubmed:29238626
  • Influence of micro-and macro-vascular disease and Tumor Necrosis Factor Receptor 1 on the level of lower-extremity amputation in patients with type 2 …Pubmed:29879997
  • Circulating angiopoietin‑like protein 2 levels are associated with decreased renal function in HIV+ subjects on cART: A potential marker of kidney disease
  • Serum tenascin-C is independently associated with increased major adverse cardiovascular events and death in individuals with type 2 diabetes: a French …Pubmed: 32040670

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