Application Note
LCAT (Phosphatidylcholine-sterol acyltransferase) is an enzyme in the extracellular metabolism of plasma lipoproteins, which converts cholesterol and phosphatidylcholines (lecithins) to cholesteryl esters and lysophosphatidylcholines. It is reported that APOA1 (Apolipoprotein A-I) acts as a cofactor for the LCAT. Thus, a binding ELISA assay was conducted to detect the association of LCAT with APOA1. Briefly, recombinant rat LCAT were diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to APOA1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-LCAT pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of LCAT with APOA1 was in a dose dependent manner.
Observed MW
22 kDa.
Form
Lyophilized powder
Buffer
Reconstitute with 10mM PBS (pH7.4) to 0.1-1.0mg/ml. Do not vortex. Lyophilized from PBS (pH7.4), 0.01% SKL, 1mM DTT, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Glu41~His210 (NP_032516.2)
Expression System
E. coli
Purity
> 90%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.