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Active Melanoma Inhibitory Activity Protein 1 (MIA1)

CD-RAP; Melanoma-derived growth regulatory protein

ACTIVITY TEST

Melanoma inhibitory activity (MIA), also known as cartilage-derived retinoic acid-sensitive protein (CD-RAP), is a 12-kDa protein that is secreted from both chondrocytes and malignant melanoma cells. MIA has been reported to have effects on cell growth and adhesion, and it may play a role in melanoma metastasis and cartilage development. In the presence of MIA, we observed enhanced migratory ability of melanocytic cells, induction of melanoma-associated genes as well as inhibition of apoptosis due to anoikis. S100 Calcium Binding Protein B (S100B) is a high affinity ligand for MIA1, a functional binding ELISA assay was conducted to detect the interaction of recombinant human MIA1 and recombinant bovine S100B. Briefly, MIA1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to S100B-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MIA1 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 450/630 nm immediately. The binding activity of recombinant humant MIA1 and recombinant bovine S100B was shown in Figure 1, the EC50 for this effect is 1.1 ug/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) 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 Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
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