Human Apolipoprotein E/ApoE Quantikine ELISA Kit

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Human Apolipoprotein E/ApoE Standard Curve
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Product Details
Procedure
Citations (2)
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Human Apolipoprotein E/ApoE Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (50 uL), Urine (50 uL), Human Milk (10 uL)
Sensitivity
0.709 ng/mL
Assay Range
3.1 - 200 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
Specificity
Natural and recombinant human Total ApoE (ApoE2, ApoE3, and ApoE4).
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Apolipoprotein E/ApoE Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human ApoE in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains E. coli-expressed recombinant human ApoE and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human ApoE showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human ApoE.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 29.1 59.6 118 28.1 60.6 119
Standard Deviation 0.913 1.38 2.91 1.71 4.17 3.89
CV% 3.1 2.3 2.5 6.1 6.9 3.3

Recovery

The recovery of human ApoE spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 105 93-110

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human ApoE were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Apolipoprotein E/ApoE ELISA Linearity

Data Examples

Human Apolipoprotein E/ApoE Standard Curve

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Apolipoprotein E/ApoE

The apolipoproteins are a structurally-unrelated group of proteins that have some association with the transport of lipids in blood. Apolipoproteins, plus phospholipids, cholesterol and triglycerides, form spherical particles with a lipid/hydrophobic center and a (apolipo)protein coat. The apolipoprotein coat promotes aqueous solubility and serves as a ligand for lipoprotein receptors. HDL may contain apolipoproteins A, C, D, E, J, L and M, while LDL contains apolipoproteins B and E.

ApoAI and ApoA2 are major protein components of serum high-density lipoprotein (HDL) and are produced by the liver and small intestine. They are involved in reverse cholesterol transport from tissues to the liver. Polymorphisms of ApoA2 are associated with disorders of cholesterol and fatty acid metabolism. Human ApoB (Apolipoprotein B-100) is a 550 kDa, secreted, palmitoylated glycoprotein that is part of LDL and VLDL particles. It is made by liver and is 4536 aa in length. It binds LDL to the ApoB/E receptor. ApoC activates lipoprotein lipase and may self-associate to form amyloid-type fibrils.

ApoE is a 34 kDa protein component of serum chylomicrons, VLDL, and HDL particles. It mediates the binding, uptake, and catabolism of these particles through interactions with the ApoE receptor and LDL receptors in the liver and brain. ApoE is important in fatty acid homeostasis and memory formation. Polymorphisms encode three variants (ApoE2, 3, 4) which are differentially related to the development of atherosclerosis and neurogenerative disorders, particularly Alzheimer's disease.

Serum amyloid A proteins (SAAs) are a family of homologous apolipoproteins of high density lipoprotein (HDL). They can be divided into two groups. The first group consists of the acute phase SAA1 and SAA2 that associate with HDL during inflammation and remodel the HDL particle by displacing apolipoprotein A1. The second group consists of constitutively expressed SAA4 and SAA5 that exist as minor apolipoproteins on HDL but make up more than 90% of the total SAA during homeostasis.

Entrez Gene IDs:
348 (Human); 11816 (Mouse); 25728 (Rat)
Alternate Names:
APOE; Apo-E; Apolipoprotein E; apolipoprotein E3; late onset); MGC1571
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Apolipoprotein E/ApoE Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Apolipoprotein E2 and E3, but Not E4, Promote Retinal Pathologic Neovascularization
    Authors: T Masuda, M Shimazawa, Y Hashimoto, A Kojima, S Nakamura, S Suemori, K Mochizuki, H Kawakami, K Kawase, H Hara
    Invest. Ophthalmol. Vis. Sci., 2017;58(2):1208-1217.
    Species: Human
    Sample Types: Vitreous Humor
  2. Biomarkers of inflammation and innate immunity in atrophic nonunion fracture
    J Transl Med, 2016;14(0):258.
    Species: Human
    Sample Types: Serum

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Human Apolipoprotein E/ApoE Quantikine ELISA Kit
By Anonymous on 06/21/2016
Sample Tested: Serum