ELISA Kit for Apolipoprotein E (APOE)

Apo-E; AD2; Apoprotein; Alzheimer Disease 2(E4-Associated,Late Onset

Specificity

This assay has high sensitivity and excellent specificity for detection of Apolipoprotein E (APOE).
No significant cross-reactivity or interference between Apolipoprotein E (APOE) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Apolipoprotein E (APOE) and the recovery rates were calculated by comparing the measured value to the expected amount of Apolipoprotein E (APOE) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-105 88
EDTA plasma(n=5) 90-104 101
heparin plasma(n=5) 99-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apolipoprotein E (APOE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apolipoprotein E (APOE) 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 Apolipoprotein E (APOE) 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) 94-103% 86-95% 95-104% 91-105%
EDTA plasma(n=5) 84-101% 88-102% 95-102% 90-97%
heparin plasma(n=5) 98-105% 80-96% 85-98% 81-102%

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.

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Magazine Citations
Seizure - European Journal of Epilepsy Cerebrospinal fluid apolipoprotein E concentration decreases after seizure Seizure: PIIS1059131109002441
Biochimica et Biophysica Acta Hepatic lipase- and endothelial lipase-deficiency in mice promotes macrophage-to-feces RCT and HDL antioxidant properties PubMed: 23328279
Medical Oncology Expression and clinical significance of apolipoprotein E in pancreatic ductal adenocarcinoma Pubmed: 23609192
Diabetes. Adipocyte Atp-Binding Cassette G1 Promotes Triglyceride Storage, Fat Mass Growth And Human Obesity Pubmed:25249572
renal failure Frequencies of apolipoprotein E alleles in depressed patients undergoing hemodialysis – a case-control study PubMed: 25707516
Biochim Biophys Acta Culturing of HepG2 cells with human serum improve their functionality and suitability in studies of lipid metabolism PubMed: 26515253
Diabetes Adipocyte ATP-binding cassette G1 promotes triglyceride storage, fat mass growth, and human obesity PubMed: 25249572
Scientific Reports Adaptive immunity against gut microbiota enhances apoE-mediated immune regulation andreduces atherosclerosis and western-diet-related inflammation. pubmed:27383250
Biochemical Pharmacology  Differential effects of apoE and apoJ mimetic peptides on the action of an anti-Aβ scFv in 3xTg-AD mice Pubmed:30026023
Oncotarget Deletion of plasma Phospholipid Transfer Protein (PLTP) increases microglial phagocytosis and reduces cerebral amyloid-β deposition in the J20 mouse … Pubmed:29731975
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