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ACTIVITY TEST
Buffer Formulation 20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300. Traits Freeze-dried powder Purity > 90% Isoelectric Point 6.2 Applications Cell culture; Activity Assays.
Fibronectin type III domain-containing protein 5, the precursor of irisin, is a protein that is encoded by the FNDC5 gene. It was reported that FNDC5 significantly decreased cell number, migration and viability through apoptosis in malignant MDA-MB-231 cells. Thus MDA-MB-231 cells were seeded overnight at a density of 5,000 cells/well, and treated with or without various concentrations of FNDC5 for 48h, then MDA-MB-231 cells were observed by inverted microscope and cell viability was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10µL of CCK-8 solution was added to each well of the plate, then measure the absorbance at 450nm using a microplate reader after incubating the plate for 1-4 hours at 37℃.
Cell apoptosis of MDA-MB-231 cells after incubation with FNDC5 for 48h observed by inverted microscope was shown in Figure 1.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.
Image
Figure. SDS-PAGE
Figure. Western Blot
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