| Product Description | SpeciesGene IDAccessionSourceLengthMWTagHuman7448P04004E. coli~400aa46kDa6X HisAbout this proteinNameUltraBio™ Human Vitronectin (Embryo cell culture grade); UltraBio™ Human Vitronectin (Embryo cell culture tested); Recombinant Human VitronectinSynonymsVN; V75; VNT; VTN-N; rh-VTN; rhVTN; Vitronectin; S-Protein; Serum-Spreading Factor; EpibolinSourceRecombinant truncated human vitronectin (62-478aa) purified from E. coli.PuritySterile, ≥ 95% by SDS-PAGEBiological ActivityUsed as a substrate for the feeder-free culture of human pluripotent stem cells (PSCs) and has been proven to maintain pluripotency and normal growth characteristics in multiple PSC lines such as H9 human embryonic stem cells. Measured by the ability of the immobilized protein to support the adhesion of DU145 human prostate carcinoma cells. When cells are added to Vitronectin-coated plates (10μg/ml and 100μl/well), approximately ＞40% cells will adhere specifically in 30 minutes at 37℃. Physical AppearanceWhite lyophilized (freeze-dried) powder.FormulationLyophilized in PBS, pH7.3±0.1.ReconstitutionWe recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in ultrapure water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0mg/ml. Stock solutions should be apportioned into working aliquots and stored at ≤ -20ºC. Further dilutions should be made in appropriate buffered solutions.CategoryExtracellular Matrix (ECM), Cell Culture, Embryo cell cultureBackgroundVitronectin has well-defined activity in cell adhesion, thrombosis, complement activation, fibrinolysis, inflammation, and platelet adhesion. Vitronectin binds to a variety of macromolecules and participates in regulatory circuits to control these processes. Some target macromolecules that interact with vitronectin are heparin, PAI-1, proteases such as thrombin and urokinase-type plasminogen activator (uPA), serine protease inhibitor-protease complexes, the uPA receptor, and a sub-class of integrin receptors on the surface of cells. Vitronectin is also applied as a thin coating on tissue-culture surfaces, which is useful to promote cell attachment, spreading, proliferation, and differentiation of many normal and neoplastic cells, and to study cell migration [1,2].Store at -20℃, valid for at least 1 year.Please refer to Figure 1 for the SDS-PAGE analysis of UltraBio™ Human Vitronectin.Figure 1. SDS-PAGE analysis of Aladdin's UltraBio™ Human Vitronectin with UltraBio™ Plus PAGE Precast Gel (Tris-Gly, 12%, 15-well) . Prestained Protein Ladder (10-170kDa) was used as a marker and the gel was stained with UltraBio™ Komas Brilliant Blue Ultrafast Staining Solution . This figure is for reference only, which may vary due to different experimental conditions.This product is provided in powder. A sterile 1mg/ml stock solution can be prepared by dissolving this product with an appropriate amount of cell culture grade ultrapure water , and then be diluted appropriately for coating cell culture vessels.This product can be used for normal culture and stemness maintenance of human embryonic stem cells or multipotential stem cells under feeder layer cell-free conditions. Please refer to Figure 2 for the morphology and growth of H9 human embryonic stem cells cultured with this product.Figure 2.Morphology and growth of H9 human embryonic stem cells cultured with Aladdin's UltraBio™ Human Vitronectin and Competitor from T Company. As shown in the figure, this product has similar performance to the Competitor in maintaining and promoting the growth of H9 human embryonic stem cells. The Scale Bar is 50µm. This figure is for reference only, which may vary due to different experimental conditions.Please refer to Figure 3 for the immunofluorescence staining of H9 human embryonic stem cells cultured with this product.Figure 3.The immunofluorescence staining results of H9 human embryonic stem cells cultured with Aladdin's UltraBio™ Human Vitronectin and Competitor from T Company. Aladdin's Pluripotent Stem Cell Immunofluorescence Detection Kit (Oct4, red fluorescence) and Pluripotent Stem Cell Immunofluorescence Detection Kit (Sox2, red fluorescence) were used for immunofluorescence staining. As shown in the figure, this product has similar performance to the Competitor in maintaining and promoting the growth of H9 human embryonic stem cells. The Scale Bar is 50µm. This figure is for reference only, which may vary due to different experimental conditions.Please refer to Figure 4 for the viability assay of H9 human embryonic stem cells cultured with this product.Figure 4.Cell viability assay of H9 human embryonic stem cells cultured with Aladdin's UltraBio™ Human Vitronectin and Competitor from T Company. Cell viability was assayed using CCK-8 . As shown in the figure, this product has similar performance to the Competitor in maintaining and promoting the growth of H9 human embryonic stem cells. This figure is for reference only, which may vary due to different experimental conditions.
Precautions： The human vitronectin solutions should be sterilized by filtration and handled aseptically to avoid microbial contamination of cell cultures.To avoid non-specific adsorption of trace proteins to the surface of centrifuge tubes, we recommend preparing the stock solution with a solution containing an appropriate amount of BSA.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.
Instructions for Use： 1.Immediately store this product at -20℃ upon receipt. Before opening the cap for the first time, centrifuge at 8,000-12,000×g for approximately 10 seconds to collect the powder at the bottom of the tube.2.Prepare a stock solution at 1mg/ml by dissolving this product with an appropriate amount of cell culture grade ultrapure water (, ST875), and sterilize the stock solution by filtration. Prepare an appropriate amount of working solution by diluting the stock solution as required by particular experiments. Do not shake the human vitronectin solutions vigorously to avoid protein denaturation and inactivation.3.Please refer to relevant literature for an optimal working concentration of human vitronectin for a particular experiment, or optimize through tests. The commonly used dosage of human vitronectin is 0.5-5μg/cm2.4.Cell culture plates to be used for H9 human embryonic stem cells can be coated as follows:a) Before opening the cap of this product for the first time, centrifuge at 8,000-12,000×g for approximately 10 seconds to collect the powder at the bottom of the tube.b) Prepare a stock solution at 1mg/ml by dissolving this product with an appropriate amount of cell culture grade ultrapure water (, ST875), and sterilize the stock solution by filtration. Aliquot 20μl into each 1.5ml centrifuge tube and store at -20ºC.c) Dilution: Thaw human vitronectin (1mg/ml) at room temperature. Dilute it to a final concentration of 30μg/ml using 1X PBS (, C0221A) and mix gently. Extra working solutions can be frozen in aliquot to avoid repeated freeze-thaw.d) Coating: Add an appropriate volume of human vitronectin working solution (30μg/ml) to cell culture plates at 3μg/cm2, as recommended in the table below. Shake the plates gently to cover the bottom of the plate evenly, then incubate at 37℃ for 2h.Plates or DishesGrowth AreaVolume12-well plate3.8 cm2/well0.38ml/well6-well plate9.5 cm2/well0.95ml/well3.5mm dish9 cm2/dish0.9ml/dish60mm dish21 cm2/dish2.1ml/dish100mm dish55 cm2/dish5.5ml/dishNote 1: Cell culture plates need to be sealed with sealing film during incubation to avoid drying of human vitronectin working solution.Note 2: If not used immediately, the coated plates can be stored at 4℃ for up to one week and be equilibrated at room temperature for 30 minutes before use.e) Discard the human vitronectin working solution and seed cells after adding an appropriate volume of medium.
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