Homocysteine Assay Kit (Fluorometric)

AgK-421

Homocysteine Assay Kit (Fluorometric)

Homocysteine Assay Kit allows for quantification of total homocysteine in biological fluids such as plasma and serum. The assay is based on the reduction of homocysteine disulfides to free homocysteine, which is cleaved by a homocysteine-selective enzyme, generating an intermediate product. The intermediate reacts with a probe solution to form a stable fluorophore that emits in the far-red spectrum (Ex/Em = 658/708 nm). The assay is not affected by physiological concentrations of other biological thiols (such as cysteine, methionine and glutathione), is high-throughput adaptable and can detect as low as 5 μM homocysteine.

Homocysteine is a non-proteogenic amino acid synthesized intracellularly by removal of the N-methyl group from the essential amino acid methionine. Homocysteine is exported from cells into the blood, where it exists mainly as an oxidized disulfide species, either as a dimer or bound to cysteine residues of serum proteins. The reduced form of homocysteine (‘free’ homocysteine) can be metabolized into cysteine via the transsulfuration pathway; however, it can also undergo intramolecular cyclization, forming the highly reactive pro-oxidant homocysteine thiolactone. Subsequent N-homocysteinylation of protein lysine residues by the reactive thiolactone disrupts protein conformation, leading to formation of cytotoxic protein aggregates. Homocysteinylated proteins may also act as autoantigens, triggering arterial inflammation and atherosclerosis. Elevated plasma homocysteine concentration is a clinical biomarker for increased risk of cardiovascular disease, ischemic stroke and myocardial infarction. Severely elevated homocysteine levels (hyperhomocysteinemia) are correlated with a 4-fold increase in mortality due to heart attack and a 16-fold increase in the likelihood of recurrent stroke.

Estimation of Homocysteine in various biological samples

Store at -20°C.

100 assays

• Homocysteine Assay Buffer
• Disulfide Reducing Agent (DTT)
• Homocysteine Enzyme Mix
• Fluorogenic Probe Solution
• Develop Solution
• Homocysteine Disulfide Standard

Multiwell fluorescence microplate reader
Precision multi-channel pipette and reagent reservoir
White 96-well plates with flat bottom

Fluorescence (Ex/Em = 658/708 nm)

Human or animal plasma or serum