Human Apolipoprotein E4 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Apolipoprotein E4 in Serum, Citrate plasma, EDTA Plasma, Heparin Plasma, Cell culture supernatant samples.
APOE is an apolipoprotein, a protein associating with lipid particles, that mainly functions in lipoprotein-mediated lipid transport between organs via the plasma and interstitial fluids (PubMed:14754908, PubMed:1911868, PubMed:6860692). APOE is a core component of plasma lipoproteins and is involved in their production, conversion and clearance (PubMed:14754908, PubMed:1911868, PubMed:1917954, PubMed:23620513, PubMed:2762297, PubMed:6860692, PubMed:9395455). Apolipoproteins are amphipathic molecules that interact both with lipids of the lipoprotein particle core and the aqueous environment of the plasma (PubMed:2762297, PubMed:6860692, PubMed:9395455). As such, APOE associates with chylomicrons, chylomicron remnants, very low density lipoproteins (VLDL) and intermediate density lipoproteins (IDL) but shows a preferential binding to high-density lipoproteins (HDL) (PubMed:1911868, PubMed:6860692). It also binds a wide range of cellular receptors including the LDL receptor/LDLR, the LDL receptor-related proteins LRP1, LRP2 and LRP8 and the very low-density lipoprotein receptor/VLDLR that mediate the cellular uptake of the APOE-containing lipoprotein particles (PubMed:12950167, PubMed:1530612, PubMed:1917954, PubMed:20030366, PubMed:20303980, PubMed:2063194, PubMed:2762297, PubMed:7635945, PubMed:7768901, PubMed:8756331, PubMed:8939961). Finally, APOE has also a heparin-binding activity and binds heparan-sulfate proteoglycans on the surface of cells, a property that supports the capture and the receptor-mediated uptake of APOE-containing lipoproteins by cells (PubMed:23676495, PubMed:7635945, PubMed:9395455, PubMed:9488694). A main function of APOE is to mediate lipoprotein clearance through the uptake of chylomicrons, VLDLs, and HDLs by hepatocytes (PubMed:1911868, PubMed:1917954, PubMed:23676495, PubMed:29516132, PubMed:9395455). APOE is also involved in the biosynthesis by the liver of VLDLs as well as their uptake by peripheral tissues ensuring the delivery of triglycerides and energy storage in muscle, heart and adipose tissues (PubMed:2762297, PubMed:29516132). By participating in the lipoprotein-mediated distribution of lipids among tissues, APOE plays a critical role in plasma and tissues lipid homeostasis (PubMed:1917954, PubMed:2762297, PubMed:29516132). APOE is also involved in two steps of reverse cholesterol transport, the HDLs-mediated transport of cholesterol from peripheral tissues to the liver, and thereby plays an important role in cholesterol homeostasis (PubMed:14754908, PubMed:23620513, PubMed:9395455). First, it is functionally associated with ABCA1 in the biogenesis of HDLs in tissues (PubMed:14754908, PubMed:23620513). Second, it is enriched in circulating HDLs and mediates their uptake by hepatocytes (PubMed:9395455). APOE also plays an important role in lipid transport in the central nervous system, regulating neuron survival and sprouting (PubMed:25173806, PubMed:8939961). APOE is also involved in innate and adaptive immune responses, controlling for instance the survival of myeloid-derived suppressor cells (By similarity). Binds to the immune cell receptor LILRB4 (PubMed:30333625). APOE may also play a role in transcription regulation through a receptor-dependent and cholesterol-independent mechanism, that activates MAP3K12 and a non-canonical MAPK signal transduction pathway that results in enhanced AP-1-mediated transcription of APP (PubMed:28111074). (Microbial infection) Through its interaction with HCV envelope glycoprotein E2, participates in the attachment of HCV to HSPGs and other receptors (LDLr, VLDLr, and SR-B1) on the cell surface and to the assembly, maturation and infectivity of HCV viral particles (PubMed:25122793, PubMed:29695434). This interaction is probably promoted via the up-regulation of cellular autophagy by the virus (PubMed:29695434).
Apolipoprotein E, Apo-E, APOE
Human Apolipoprotein E4 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Apolipoprotein E4 in Serum, Citrate plasma, EDTA Plasma, Heparin Plasma, Cell culture supernatant samples.
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Serum | n 8 | mean - | SD - | C.V. 2.5 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Serum | n 3 | mean - | SD - | C.V. 4 |
Sample type | Average % | Range |
---|---|---|
Sample type Serum | Average % = 106 | Range 105 - 107 % |
Sample type Citrate plasma | Average % = 100 | Range 83 - 110 % |
Sample type EDTA Plasma | Average % = 91 | Range 84 - 101 % |
Sample type Heparin Plasma | Average % = 105 | Range 85 - 116 % |
Sample type Cell culture supernatant | Average % = 102 | Range 92 - 110 % |
Human Apolipoprotein E4 SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Apolipoprotein E4 protein in Serum, Cit Plasma, EDTA Plasma, Hep Plasma, Cell culture supernatant. Quantitate Human Apolipoprotein E4 with 366.056 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips, available in 10-pack (10 x 96-well plates)
-Also available in a fully validated 384-well format.
Apolipoprotein E4 often called ApoE4 is a member of the apolipoprotein family and plays an important role in lipid metabolism. As a 34-kDa protein it associates with lipoprotein particles and mediates receptor binding and clearance of these complexes. ApoE4 expresses mainly in the liver and the brain where it facilitates cholesterol and lipid transport and homeostasis. Other important alternative names for this protein include apo E4 and E4 peptide which identify specific functions related to its diverse roles in the body.
ApoE4 is an important player in lipid transport and uptake facilitating the redistribution of lipids among cells. It forms part of a critical complex involved in binding to low-density lipoprotein receptors (LDLR) and the related protein family enabling efficient lipid delivery to peripheral tissues. The interaction of ApoE4 with these receptors directly influences cellular lipid balance impacting important cellular functions. Additionally its polymorphic nature allows the isoforms E2 E3 and E4 to differ in their impacts on lipid transport and disease risk.
ApoE4 engages in the lipid and cholesterol transport pathways prominently impacting the reverse cholesterol transport process and the regulation of plasma lipoprotein metabolism. It interacts with proteins such as LDLR and LRP1 influencing the removal of triglyceride-rich lipoproteins and modulating lipid levels in the circulation. ApoE4's involvement in these pathways highlights its role as a central regulator of lipid homeostasis.
ApoE4 is closely associated with Alzheimer’s disease and cardiovascular disease highlighting its significance beyond lipid transport. ApoE4's isoform uniquely elevates the risk for Alzheimer's disease by potentially altering amyloid-beta metabolism and interfering with tau-associated pathologies making it a target of interest in neurodegenerative research. Additionally individuals bearing the ApoE4 allele exhibit an increased susceptibility to cardiovascular disorders due to its impact on cholesterol transport and clearance often linking it with elevated blood cholesterol levels. Through these diseases ApoE4 connects with other proteins like amyloid precursor protein (APP) and tau in Alzheimer's disease illustrating the broader implications of its biological functions.
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Interpolated concentrations of human Apolipoprotein E4 in Human Serum in 96-well vs. 384-well plates.
Interpolated concentration of native Apolipoprotein E4 was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 1:3,200 human serum. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Example of human Apolipoprotein E4 standard curve in Sample Diluent NS in 96-well vs. 384-well plate.
Example of human Apolipoprotein E4 standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
Interpolated concentrations of Apolipoprotein E4 in normal human serum donors.
Serum of eight individual healthy human male donors was measured in duplicate. Interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). The mean Apolipoprotein E4 concentration was determined to be 930.1 µg/mL with a range of 33.7 – 6,365.6 µg/mL.
Interpolated concentrations of human Apolipoprotein E4 in HepG2 cell culture supernatant.
Interpolated concentration of native Apolipoprotein E4 was measured in duplicate at different sample concentrations. Undiluted samples are as follows: HepG2 Cell Culture Supernatant 1:8. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Interpolated concentrations of human Apolipoprotein E4 in human serum, plasma – citrate, plasma – EDTA, and plasma – heparin.
Interpolated concentration of native Apolipoprotein E4 was measured in duplicate at different sample concentrations. Undiluted samples are as follows: human serum 1:3,200, human plasma – citrate 1:800, human plasma – EDTA 1:1,333, and human plasma – heparin 1:167. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Example of human Apolipoprotein E4 standard curve in Sample Diluent NS.
Example of human Apolipoprotein E4 standard curve. Background-subtracted data values (mean +/- SD) are graphed.
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