Active Vitronectin (VTN)

V75; VN; VNT; Serum Spreading Factor; Somatomedin B; Complement S-Protein

ACTIVITY TEST

Vitronectin (VTN ) is a glycoprotein of the hemopexin family which is abundantly found in serum, the extracellular matrix and bone. It is a secreted protein and exists in either a single chain form or a clipped, two chain form held together by a disulfide bond. VTN can binds to integrin alpha-V beta-3 and thus promotes cell adhesion and spreading. It also binds to plasminogen activator inhibitor-1 (PAI-1) and serves to regulate proteolysis initiated by plasminogen activation. Besides, Insulin Like Growth Factor Binding Protein 5 (IGFBP5) has been identified as an interactor of VTN, thus a binding ELISA assay was conducted to detect the interaction of recombinant Rat VTN and recombinant Rat IGFBP5. Briefly, VTN were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100ul were then transferred to IGFBP5-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-VTN 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 VTN and IGFBP5 was shown in Figure 1, and this effect was in a dose dependent manner.

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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Magazine Citations
Biomedical Engineering and Informatics Application of iTRAQ quantitative proteomics in identification of serum biomarkers in breast cancer IEEE: 6098563
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Analytical Biochemistry A multianalytical approach to investigate the effect of nanofiltration on plasma-derived factor IX clinical lots Pubmed:29154788
Journal of Materials Chemistry B Serum protein adsorption profile on BCP ceramic and influence of the elevated adsorption of adhesive proteins on the behaviour of MSCs Doi: 10.1039/C8TB02283F 
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