Description

Nori Porcine ApoAI ELISA Kit Summary

Alternative names for ApoAI: Apolipoprotein A-I, ApoA1,

Alternative name for porcine: pig

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 A-I is the major protein component of high-density lipoprotein (HDL) in plasma. Chylomicrons secreted from the intestinal enterocyte also contain ApoA1 but it is quickly transferred to HDL in the bloodstream (1). The protein promotes cholesterol efflux from tissues to the liver for excretion. It is a cofactor for lecithin cholesterol acyltransferase (LCAT) which is responsible for the formation of most plasma cholesteryl esters. ApoA-I was also isolated as a prostacyclin (PGI2) stabilizing factor, and thus may have an anticlotting effect (2). Defects in the gene encoding it are associated with HDL deficiencies, including Tangier disease, and with systemic non-neuropathic amyloidosis (3). A G/A polymorphism in the promoter of the ApoA-I gene has been associated with the age at which patients presented with Alzheimer disease (4). Protection from Alzheimer disease by ApoA1 may rely on a synergistic interaction with alpha-tocopherol (5). Amyloid deposited in the knee following surgery consists largely of ApoA-I secreted from chondrocytes (cartilage cells) (6). A wide variety of amyloidosis symptoms are associated with rare ApoA-I mutants. ApoA-I binds to lipopolysaccharide or endotoxin, and has a major role in the anti-endotoxin function of HDL.

References

1. Wasan KM, et al. (2008). Nature Reviews Drug Discovery 7 (1): 84
2. Yui Y, et al. (1988). J. Clin. Invest. 82 (3): 803–7.
3. Entrez Gene: APOA1 apolipoprotein A-I.
4. Vollbach H, et al. (2005). Ann. Neurol. 58 (3): 436
5. Maezawa I, at al. (2004). J. Neurochem. 91 (6): 1312
6. Solomon A, et al. (2006). Arthritis Rheum. 54 (11): 3545.

Product Citation