40 results about "Percoll" patented technology

The invention discloses a granulocyte separating medium, and a method for granulocyte separating and activity detecting by using the granulocyte separating medium. The granulocyte separating medium is prepared from percoll, a 10* phosphate buffer solution and physiological saline, wherein a volume ratio of the percoll to the 10* phosphate buffer solution to the physiological saline is 3-8:1:1-5. With the present invention, the special granulocyte separating medium is adopted to separate the granulocyte, and then detect the activity of the granulocyte so as to screen the high-performance granulocyte, improve the tumor therapy effect of the granulocyte, and easily realize targeted selection of the donor.

An optimized karyote in-vitro separation kit comprises two reagents. The reagent 1 is composed of erythrocyte precipitant, sodium chloride or potassium chloride. The reagent 2 is composed of natrii diatrizoas and polysucrose No.400, natrii diatrizoas and polysucrose dextranum, or percoll, and has density of 1.076-1.090. An application method of the optimized karyote in-vitro separation kit includes directly adding 100ml of human marrow blood, umbilical cord blood or peripheral blood into 200ml of solution of the reagent 1, allowing for standing for 10-30 minutes, collecting supernate, centrifuging at 2000rpm (1100xg) for 5 minutes, discarding the supernate, adding remnant a centrifugal tube containing solution of the reagent 2, performing density gradient centrifugation for 30 minutes, controlling the rotation speed at 1500rpm, absorbing a karyote layer, performing centrifugal cleaning for once to three times, using 50ml of cell preservative solution for diluting, and taking 100 microliters for testing or self-testing.

The invention belongs to the field of biological chemistry and specifically relates to extraction method of chloroplast protein of xylophyta ginkgo leaf. The method comprises the following steps of: firstly, preparing coarse chloroplast particles from the ginkgo leaves, performing gradient centrifugation through Percoll, so as to obtain complete chloroplast by a specific centrifugal force; then, dissolving protein by using a protein extraction buffer solution, adding the same volume of Tris-balanced phenol for extraction, and precipitating protein by using a ammonium acetate methanol solution. The extraction method is capable of effectively increase extraction efficiency and purity of chloroplast protein. The ginkgo leaf chloroplast protein obtained by the method can be used for proteomics analysis of ginkgo leaf and other xylophyta materials rich in interfering substances, so that solubility and purity of protein are improved; a 2-DE map of the protein has a clear background and many protein spots are distinguishable.

This invention provides a solution for preserving mammalian early embryos or ES cells by vitrification, which comprises, as a base material, a phosphate buffer that exclusively contains 10% to 15% (v/v) propylene glycol as polyhydric alcohol or a phosphate buffer that exclusively contains 10% to 15% (v/v) propylene glycol and 25% to 35% (v/v) ethylene glycol as polyhydric alcohols and further contains 15% to 25% (v/v) Percoll® and 0.2 M to 0.5 M sucrose. This invention also provides a method for preserving mammalian early embryos or ES cells by vitrification using such solution.

The invention belongs to the field of tumour basic research, and relates to a method used separating and purifying microglia cells from glioma specimens. The method comprises following steps: a cell suspension is prepared from tumour tissues via mechanical method combined with enzymatic digestion method; the cell suspension is subjected to density gradient centrifugation so as to obtain the microglia cells of CD11B+ with a relatively high purity via enrichment and selection; and the microglia cells with a relatively high purity are obtained via further separation using CD11B+ magnetic beads. Wherein, Percoll separating medium is composed of 70% of Percoll, 30% of Percoll, and HBSS. The purity of the microglia cells obtained via percoll density gradient centrifugation enrichment can be more than 95%, and it is confirmed by immunofluorescence assay and phagocytosis assay that functions of the microglia cells are excellent. The method is simple and convenient for operation, is low in cost, is stable in quality, and is capable of providing basis for further research on immunologic functions microglia cells in brain glioma and effects of microglia cells in immune escape of tumour.

The invention discloses a hydrogen production chlamydomonas chloroplast separation method which comprises the following steps: 1) hanging Chlamydomonas in a lysate, adding saponin until the final concentration is 0.25g / 100mL; 2) putting the step 1) solution on the ice for 1-2min for chlamydomonas cell lysis; 3) centrifuging the step 2) solution for 1min at 2300g at 4 DEG C; 4) using the lysate for rinsing steps 3) precipitate, centrifuging for 1min at 1000-2300g at 4 DEG C to obtain crude chloroplast extract; 5) hanging the crude chloroplast extract in a lysate; 6) adding 75% by volume of Percoll on the centrifugal pipe bottom, adding 45% by volume of Percoll into the upper layer, adding the step 5) solution into the uppermost layer to the form the gradient, centrifuging for 20min at 4600g at 4 DEG C to form dark green bands between two different volume fractions of Percoll; and 7) taking the dark green bands out, using the lysate to dilute, centrifuging for 2min at 890-1000g at 4 DEG C, and collecting precipitate to obtain chlamydomonas chloroplast. The experiment proves that the method has the advantages of simple operation, high chlamydomonas chloroplast yield and shorter time.

The invention discloses a method for separating and purifying protoplast of rice. The method comprises following steps: A, pretreatment in a CPW salt solution; B, enzymolysis; C, dilution and precipitation; D, centrifugation; E, resuspending. The step A of pretreatment in the CPW salt solution further comprises steps as follows: callus tissue obtained by fluid suspension culture is collected and slightly flicked, tissue blocks are enabled to be dispersed to the greatest extent, the tissue blocks are cleaned once by a culture solution and cleaned 2-3 times by the CPW salt solution, and a product is put in the CPW salt solution for pretreatment. With the adoption of the method of density gradient centrifugation based on a silica gel particle suspension (namely, a Percoll layering liquid) which is treated by polyvinylpyrrolidone, damage to cells is greatly reduced, time is shortened, protoplast cells with high activity can be obtained by separation, cell sizes are kept relatively consistent, and convenience is brought to follow-up transformation and biological operation of other molecules.

The invention relates to a method for separating and purifying CD8+T cells from liver cancer tissue. The method for separating the CD8+T cells comprises the following specific steps: (1) separating tumor infiltration immune cells; (2) performing discontinuous density gradient centrifugation with 30% percoll and 60% percoll; and (3) sorting the CD8+T cells by using MACS (Magnetic Activated Cell Sorting) microbeads. With the combination of density gradient centrifugation and microbead sorting, the CD8+T cells are successfully purified from liver cancer tissue, the purity of the cells is identified by using a flow cytometry, the purity is up to 95% or greater, and meanwhile the cell number is up to 3*10<5> or higher.

The invention relates to an application of percoll to improving separation efficiency of a sperm flow cytometer, belongs to the technical field of animal breeding, and is used for improving the separation efficiency of sperm flow cells. The application disclosed by the invention has the advantages that by a percoll method, the vitality of sperms can be notably improved, dead sperms can be removed,and the separation efficiency of the sperm flow cells can be effectively improved through treated sperms.

The invention provides a separation method for separating peripheral blood mononuclear cells, and particularly relates to the field of biological medicines. The separation method comprises the following steps: S1, preparation of a separation tube: firstly, adding 2-6 ml of Percoll or polysucrose or meglumine diatrizoate cell separation liquid with the density of 1.075-1.0796 g/ml into a centrifugal tube, sucking 0.5-1.5 ml of separation gel with the density of 1.06-1.07 g/ml, adding the separation gel into a tube opening of the centrifugal tube, and carrying out horizontal centrifuging for 1-3minutes at room temperature under the centrifugal force of 800-1200 g, so that the separation gel forms an isolation layer on the liquid surface of the Percoll or polysucrose or meglumine diatrizoatecell separation liquid, and preparation of the separation tube is finished; and S2, separation of the peripheral blood mononuclear cells: adding 2-6 ml of anticoagulant whole blood into the preparedseparation tube, and carrying out horizontal centrifuging for 8-12 minutes at room temperature under the centrifugal force of 800-1200 g; sucking and discarding the uppermost liquid to remove cell fragments and platelets, directly pouring the liquid above the isolation layer into a collection tube, carrying out centrifuging for 4-6 minutes at room temperature under the centrifugal force of 600-1000 g, and resuspending the cells by using PBS to obtain the peripheral blood mononuclear cells. According to the separation method, it can be ensured that the anticoagulant whole blood can be absolutely not mixed with a cell separation medium after being added, and other cells are not polluted when the peripheral blood mononuclear cells are harvested.

The present invention discloses a separating and culturing method for feeder-layer-free rat spermatogonial stem cells. The spermatogonial stem cells are separated from testicles of suckling male ratsin 7-9 days after birth, differential adherence and percoll separation are conducted to obtain the rat spermatogonial stem cells with a higher purity, the obtained rat spermatogonial stem cells are cultured in a trophoblast-free culture system, can be stably passaged for a long time and can differentiate into spermatogenesis. The separating and culturing method is low in cost and simple in operation, the number of the obtained spermatogonial stem cells can be doubled, and the separating and culturing method is of great significance for treatment of infertility and protection of endangered species resources.

The invention provides a novel granulocyte separation method. The novel granulocyte separation method specifically comprises the following steps of: preparing percoll with the density of 1.090 g/mL; performing gradient density centrifugation on peripheral blood by using ficoll with the commercialized density of 1.077 g/mL; removing a supernatant, a middle albuginea layer and a ficoll layer, and leaving a lowermost erythrocyte mixture; cleaning the mixture once by using a PBS solution, and removing ficoll in the mixture; slowly adding the washed cell mixture into prepared percoll with the density of 1.090 g/mL, and carrying out centrifugation on 400g of the mixture for 20 minutes; taking out the middle layer after centrifugation is finished; carrying out splitting decomposition treatment on erythrocytes mixed in the middle layer after centrifugation by using an erythrocyte splitting decomposition solution; and adding a PBS, and washing off erythrocyte lysate to obtain the granulocyte. According to the invention, the purity of granulocytes can be greatly improved; and compared with a magnetic bead sorting method, the cost is reduced.

The invention discloses a mouse bone marrow dendritic cell induction and purification method which comprises the following steps: S1, mouse bone marrow cell extraction: clamping and cracking reservedends of thighbone and tibia by using surgical forceps, and continuously performing washing to obtain cells to the maximum extent; S2, dendritic cell culture: keeping an original culture medium and cells on a third day of culture, and adding a fresh culture medium for continuous culture; S3, purification of dendritic cell cells: adopting Percoll stratified liquid for purification and induction. According to the invention, the epiphysis is clipped and broken by surgical forceps instead of being cut off, traditional cells discarded after centrifugation on the third day are added into a culture dish again, and an original culture medium is reserved on the basis of adding the fresh culture medium during liquid change on the third day, so that the DC number of harvested cells is greatly increased; according to the method, the purity of the obtained DC cells reaches 90% or above through Percoll liquid purification and screening, the yield is increased by 21% compared with a classic magnetic bead sorting mode, and the Percoll density sorting method is lower in cost consumption.

The present invention discloses a density gradient liquid kit for myocardial cell isolation and purification and a use method thereof, and relates to the technical field of biological cell culture. The kit includes a first reagent and a second reagent. The first reagent includes ultra pure water, Percoll liquid and a buffer solution in the volume ratio of 1:3:1; the second reagent includes ultra the pure water, the Percoll liquid and the buffer solution in the volume ratio of 2:2:1; the buffer solution of the first reagent and the second reagent includes 35.064-40.032 parts of NaCl, 1.975-2.013 parts of KCl, 0.615-1.107 parts of MgSO4 .7H2O, 4.865-5.226 parts of glucose, 0.301-0.384 parts of KH2PO4, 0.213-0.365 parts of Na2HPO4 and 23.830-29.788 parts of 4-hydroxyethylpiperazine ethane sulfonic acid.