Human Delta-aminolevulinic acid dehydratase (ALAD) ELISA Kit

The ALAD enzyme is composed of 8 identical subunits and catalyzes the condensation of 2 molecules of delta-aminolevulinate to form porphobilinogen (a precursor of heme, cytochromes and other hemoproteins) . ALAD catalyzes the second step in the porphyrin and heme biosynthetic pathway; zinc is essential for enzymatic activity. ALAD enzymatic activity is inhibited by lead and a defect in the ALAD structural gene can cause increased sensitivity to lead poisoning and acute hepatic porphyria. Alternatively spliced transcript variants encoding different isoforms have been identified.

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Human Delta-aminolevulinic acid dehydratase (ALAD) ELISA Kit has high sensitivity and excellent specificity for detection of Human ALAD. No significant cross-reactivity or interference between Human ALAD and analogues was observed.

This Human Delta-aminolevulinic acid dehydratase (ALAD) ELISA Kit employs a two-site sandwich ELISA to quantitate ALAD in samples. An antibody specific for ALAD has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyALAD present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ALAD is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of ALAD bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.