Active Asporin (ASPN)

PLAP1; SLRR1C; Periodontal ligament-associated protein 1

ACTIVITY TEST

Figure. The binding activity of ASPN with PPIE.
Asporin (ASPN) is a protein belongs to a family of leucine-rich repeat (LRR) proteins associated with the cartilage matrix. The protein negatively regulates chondrogenesis in the articular cartilage and periodontal ligament (PDL) differentiation, inhibits BMP2-induced cytodifferentiation of PDL cells and also nhibits the interaction between TGFB1 and TGF-beta receptor type II in the presence of heparin/heparan sulfate in vitro. Besides, Peptidylprolyl Isomerase E (PPIE) has been identified as an interactor of ASPN, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse ASPN and recombinant mouse PPIE. Briefly, ASPN were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to PPIE-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-ASPN 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 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of ASPN and PPIE was shown in Figure 1, and this effect was in a dose dependent manner.

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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