Description

Nori Canine Apob ELISA Kit Summary

Alternative names for ApoB: Apolipoprotein B

Alternative names for canine: Dog

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Apolipoprotein B (ApoB) is a protein that in Canines is encoded by the APOB gene. ApoB is the primary apolipoprotein of chylomicrons, VLDL, IDL, and LDL particles, which is responsible for carrying lipids, including cholesterol, around the body to all cells within all tissues. ApoB is the primary organizing protein component of the particles and is absolutely required for the formation of these particles and the ApoB on the LDL particle acts as a ligand for LDL receptors in various cells throughout the body. High levels of ApoB are related to heart disease and are the primary driver of plaques that cause vascular disease (atherosclerosis), commonly first becoming obviously symptomatic as heart disease, stroke & many other body wide complications after decades of progression. There is considerable evidence that concentrations of ApoB^[1] and especially the NMR assay^[2] are superior indicators of vascular/heart disease driving physiology than either total cholesterol or LDL-cholesterol. ApoB occurs in the plasma in 2 main isoforms, ApoB48 and ApoB100. The first is synthesized exclusively by the small intestine, the second by the liver.^[3] ApoB-100 is the largest of the apoB group of proteins, consisting of 4563 amino acids and both isoforms are coded by APOB.^[3] ApoB48 is generated when a stop codon (UAA) at residue 2153 is created by RNA editing. Overproduction of apolipoprotein B can result in lipid-induced endoplasmic reticulum stress and insulin resistance in the liver.^[4] ApoB100 levels are associated with coronary heart disease, the number of ApoB100-containing lipoprotein particles which can carry lipids into the artery walls is a key determinant, driver of atherosclerosis and heart disease. The ApoB100 / ApoA1 ratio is more effective at predicting heart attack risk, in patients who had had an acute myocardial infarction, than either the ApoB100 or ApoA1 measure alone.^[5] In the general population this remains unclear although in a recent study ApoB was the strongest risk marker for cardiovascular events.^[6] ApoB has been shown to interact with apo(a), PPIB,^[7] Calcitonin receptor^[7] and HSP90B1.^[7] Interaction of ApoB with proteoglycans, collagen, and fibronectin is believed to cause atherosclerosis.

References

1. Lim JS, et al. (2011). Journal of Clinical Lipidology. 5 (4): 264–272.
2. Carmena R, et al. (2004). Circulation. 109 (23): III–2.
3. Chen SH, et al. (1986). Journal of Biological Chemistry. 261 (28): 12918–12921.
4. Su Q, et al. (2009). Hepatology. 50 (1): 77–84.
5. McQueen MJ, et al. (2008). Lancet. 372 (9634): 224–33.
6. Benn M, et al. (2007). Arterioscler Thromb Vasc Biol. 27 (3): 661–70.
7. Zhang J, Herscovitz H (2003). J. Biol. Chem. 278 (9): 7459–68.

Product Citations