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Active Lipase A, Lysosomal Acid (LIPA)

LAL; CESD; Lipase A,Lysosomal Acid,Cholesterol Esterase(Wolman Disease); Acid Cholesteryl Ester Hydrolase; Lysosomal acid lipase/cholesteryl ester hydrolase

ACTIVITY TEST

Lysosomal Acid Lipase (LIPA) is a crucial enzyme within the lysosomes that hydrolyzes cholesteryl esters and triglycerides into free cholesterol and free fatty acids. This process is vital for cellular lipid metabolism and energy homeostasis. Deficiencies in LIPA activity lead to severe lipid storage disorders, such as Wolman disease and Cholesteryl Ester Storage Disease (CESD), characterized by the massive accumulation of lipids in the liver, spleen, and other organs. Regarding its interaction, LIPA does not directly bind to Apolipoprotein B (APOB); instead, it interacts with and degrades the lipid core of Low-Density Lipoprotein (LDL) particles, of which APOB is the primary structural protein component.Besides,APOB has been identified as an interactor of LIPA, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant rat LIPA and recombinant pig APOB. Briefly, LIPA was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to APOB-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-LIPA pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant rat LIPA and recombinant pig APOB was shown in Figure 1, the EC50 for this effect is 1.06326µg/mL.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
APD225Ra01 Active Lipase A, Lysosomal Acid (LIPA) Cell culture; Activity Assays.
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