From western blotting, it was verified that the SFPS 191,212 treatment improved the expression of Bax, Caspase-9, and Caspase-3 genes and proteins, while it reduced phosphatidylinositol 3 kinase and Bcl-2 genes and proteins, suggesting the involvement of mitochondria. Moreover, SPFS 191,212 increased the numbers of apoptotic cells and arrested the cell cycle in the S phase of the quantitative real-time PCR. The compound exhibited concentration-dependent effects. The anticancer activities of the SFPS 191,212 compounds were assayed in the B16F10 cells at both transcriptional and translational levels. fusiforme polysaccharides (SFPS 191,212) on the proliferation, apoptosis, and cell cycle kinetics of B16F10 murine melanoma cells were thoroughly investigated in this work. fusiforme demonstrate antitumor activities. fusiforme) is a brown alga that has been utilized as a medicine for a long time. Molecules smaller than the col- umn’s exclusion limit will be retained by the column (see Specifications).( S. Application of more or less than the recommended sample volume may decrease column per- formance.Īfter loading sample, centrifuge the column for 4 minutes at 1,000 x (g).įollowing centrifugation, the purified sample is now in either SSC or Tris buffer. Carefully apply the sample (20–75 µl) directly to the center of the column. Place the column in a clean 1.5 or 2.0 ml microcentrifuge tube. Discard the drained buffer then place the column back into the 2.0 ml tube.Ĭentrifuge for 2 minutes in a microcentrifuge at 1,000 x (g) (see Centrifugation Notes section) to remove the remain- ing packing buffer. Allow the excess packing buffer to drain by gravity to the top of the gel bed (about two minutes). If the column does not begin to flow, push the cap back on the column and then remove it again to start the flow. Snap off the tip and place the column in a 2.0 ml microcentrifuge tube (included). Invert the column sharply several times to resuspend the settled gel and remove any bubbles. PH 2–10, common aqueous buffers, formamide, dilute organic acids, alcohol, 20% (V/V) other chaotropic agents, detergents. Micro Bio-Spin columns, Bio-Gel P gel, and collection tubes are completely autoclavable at 121 ☌ for 30 minutes at pH 6.0–8.0. Microcentrifuge with a centrifugal force of 1,000 x (g).
Volumes less than 20 µl may affect recoveryīio-Gel P-6 gel: 5 base pairs (nucleic acids) or 6,000 daltons (proteins,peptides)īio-Gel P-30 gel: 20 base pairs (nucleic acids) or 40,000 dal- tons (proteins, peptides)ĩ8% retention of unincorporated nucleotides at 20 µl 90% recovery of applied DNA at 20 µlġ00% retention of unincorporated nucleotides at 20 µl 95% recovery of applied DNA at 70 µl Nucleic acids, proteins, and peptides, 20–75 µl. Tris buffer (10 mM Tris-HCl, pH 7.4) with 0.02% sodium azide SSC buffer (150 mM sodium chloride, 17.5 mM sodium cit- rate, pH 7.0) with 0.02% sodium azide
Micro Bio-Spin columns are suitable for use with 1.5 or 2.0 ml microcentrifuge tubes and are completely autoclavable.īio-Gel P-6 or P-30 polyacrylamide gel suspended in 1.0 ml of buffer This unique gel produces very effi- cient, non-interactive size separations. The columns are packed with special grades of Bio-GelĪcrylamide P-6 or P-30 gel matrices manufactured specifically for Bio-Rad spin columns. Desalt nucleic acids, proteins and peptides.Micro Bio-Spin chromatography columns are ready to use for rapid and efficient cleanup and purification of nucleic acids and proteins using a microcentrifuge.
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