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InformationAPO E ELISA KIT | FIVEphoton Biochemicals | Apolipoprotein E, Apo E | Part No. ApoE-ELISA
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Apo E (Apolipoprotein E) ELISA Kits
Part No. ApoE-ELISA.
Lead time: Approximately 2 weeks.
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Fully pre-configured sandwich ELISA format providing experimental convenience and high sensitivity.
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Strip plate configuration allowing separate use and storage of ELISA plate sections
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Affinity purified monoclonal capture antibody coated on the plate. Separate biotin-conjugated detection antibody.
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Rapid detection protocol. 1 hr total incubation at 37oC.
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Colorimetric detection.
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Suitable for CSF, ISF, cell culture medium, cell and tissue homogenates and lysates.
View Protocol |
Species |
Standard Peptide |
Assay Range |
| Human | 320 µg/ml | 1 - 300 µg/ml |
| Rat | 160 µg/ml | 50ng/ml - 150 µg/ml |
| Mouse | 96 µg/ml | 30ng/ml- 90 µg/ml |
Use the Available Options menu to select the species specificity of the ApoE ELISA kit
Overview:
ApoE transports lipoproteins, fat-soluble vitamins, and cholesterol into the lymph system and into the blood. It is synthesized in the classical secretory pathway principally in the liver and is also expressed in brain, kidney, and spleen. In the nervous system, non-neuronal cell types, most notably astroglia and microglia, are the primary producers of ApoE, while neurons express the ApoE receptors. There are seven currently identified mammalian receptors for ApoE.
ApoE has several major allelic isoforms: ApoE2, ApoE3, ApoE4: Normal ApoE-ε3: Dysfunctional ApoE-ε2 and ApoE-ε4. The major allelic forms differ by amino acid substitutions at positions 112 and 158. The E2 allele codes for Cys at positions 112 and 158 in the receptor-binding region. In contrast, the E3 allele expresses Cys-112 and Arg-158 at these positions, and the E4 allele has Arg at both positions.
E2 is associated with the genetic disorder hyperlipoproteinemia type III and atherosclerosis. E3 is found in approximately 64 percent of the population; it is considered the "neutral" Apo E genotype. E4 has been implicated in atherosclerosis and Alzheimer's disease, impaired cognitive function, and reduced neurite outgrowth.
Apo E Isolation for ELISA Assay:
Apo E undergoes synthesis, processing and secretion in the classical secretory pathway1. Therefore, it is located in compartments of the secretory pathway that include endoplasmic reticulum, Golgi and secretory vesicles, as well as the extracellular medium such as plasma and serum, and in cell culture medium. It can be isolated from intracellular compartments using detergent lysis and solubilization, or in the extracellular medium (that include plasma or cell culture medium) by sedimentation of cells and collection of the supernantant.
1. Arteriosclerosis, Thrombosis, and Vascular Biology. 2008; 28: 1060-1067
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Price (USD)
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No. of Kits
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Number of Assays |
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$645
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1
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96T |
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$1210
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2
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192T |
Methods Overview:
The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen. Samples are pre-mixed with a biotin-linked detection antibody and streptavidin-HRP, which are then added to the plate. The plate is washed after a 60 min incubation period. Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution. A stop solution is applied to terminate the reaction, and the solution then becomes yellow colored. The final reactant is measured at 450nm.
The ApoE ELISA assay kit provides a standard peptide solution with known concentration to calibrate absorbance readings to experimental sample concentration.
Image: Structure of Apo E revealing allelic variants. Source PDB:
Representative Sandwich ELISA Methods
Representative Sandwich ELISA Methods
Steps of Sandwich ELISA Assay
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Mix sample, biotinylated-detection antibody and streptavidin-HRP.
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Apply sample and antibody-HRP complex to ELISA plate for 60min at 37oC. Wash 5 times.
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Apply chromogens, incubate, and then stop the reaction.
References on ApoE
- Anchors JM, Gregg RE, Law SW, Brewer HB. ApoE deficiency: markedly decreased levels of cellular ApoE mRNA. Biochemical and Biophysical Research Communications 134: 937-943, 1986.
- Jiang Q, Lee CY, Mandrekar S, Wilkinson B, Cramer P, Zelcer N, Mann K, Lamb B, Willson TM, Collins JL, Richardson JC, Smith JD, Comery TA, Riddell D, Holtzman DM, Tontonoz P, Landreth GE. ApoE promotes the proteolytic degradation of Abeta. Neuron 58: 681-693, 2008.
- Oldfield S, Braksator E, Rodriguez-Martin I, Bailey CP, Donaldson LF, Henderson G, Kelly E. Functional interactions of APP with the apoE receptor family. Journal of Neurochemistry 106: 2263-2271, 2008.
- Rebeck GW, LaDu MJ, Estus S, Bu G, Weeber EJ. The generation and function of soluble apoE receptors in the CNS. Molecular Neurodegeneration 1: 15, 2006.
- Riddell DR, Zhou H, Atchison K, Warwick HK, Atkinson PJ, Jefferson J, Xu L, Aschmies S, Kirksey Y, Hu Y, Wagner E, Parratt A, Xu J, Li Z, Zaleska MM, Jacobsen JS, Pangalos MN, Reinhart PH. Impact of apolipoprotein E (ApoE) polymorphism on brain ApoE levels. Journal of Neuroscience 28: 11445-11453, 2008.
For research only. Not for diagnostics.
Safety: Stop solution contains acid. Avoid ingestion, skin and eye contact.
Storage: 4oC, 6 months. Strip plate sections, standard and other solutions can be stored longer term at -20oC.
Shipping. Overnight domestic delivery recommended with styrofoam box/ice pack (cost: $52 for 2 kit set). Also available for international delivery; please inquire for shipping cost.
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