细胞功能测定

1) Aliquot 5 X 106 total cells in 5-8 ml 2% media.2) Add 30 µl 3H-thymidine.3) Incubate for approximately 16 hours under normal growth conditions.4) Spin down cells and wash 2-3 times with PBS (or ser ...

1) Aliquot 5 X 106 total cells in 5-8 ml 2% media.2) Add 30 µl 3H-thymidine.3) Incubate for approximately 16 hours under normal growth conditions.4) Spin down cells and wash 2-3 times with PBS (or ser ...

death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green alive cells; P.I. (propidium iodide)-red dead cells 35 mm plates: to 2 ml med ...

death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green alive cells; P.I. (propidium iodide)-red dead cells 35 mm plates: to 2 ml med ...

Contributor: Suprya JayadevDate: December 13 19941) Transfect cells.2) Culture cells 1-3 passages in a T-75 flask containing selection material (e.g. hygromycin neomycin etc)3) Pellet cells and resusp ...

Materials: Lipofectamine Basal Medium containing 10% fetal bovine serum 1% glutamine 1% aa Basal Medium containing 1% glutamine Basal Medium containing 20% fetal bovine serum 1% glutamine 1% aa ...

Materials: Lipofectamine Basal Medium containing 10% fetal bovine serum 1% glutamine 1% aa Basal Medium containing 1% glutamine Basal Medium containing 20% fetal bovine serum 1% glutamine 1% aa ...

(Modification of methods of Dr. Seong-Seng Tan and Promega's "Protocols and Applications Guide")Cells previously transfected with a lac Z construct can be stained for b-galactosidase activity.1. Wash ...

Cultures to be treated should be sub confluent ie actively growing.1. Add 1/20 volume Mitomycin C (200 ug/ml 10 ug/ml) to culture and incubate at 37� for 2hrs.2. After approx. 2 hrs remove medium and ...

THAWING CELLSRemove a vial of Passage 2 Primary Mouse Embryo Fibroblasts (PMEF'S) at a concentration of 3 x 106 from liquid nitrogen and thaw quickly at 37oC.Add the 1ml of cell containing media to 9m ...

Area/Well Total Area Total Cells Total Vol Conc/ml Flask cm2 /cm2 5x104/cm2 96 Well 0.35 37 4.62x105 19.5 2.4X104 24 Well 1.77 42.4 5.3x105 25 2.1X104 12 Well 3.5 41.6 5.2x105 25 1.9X104 6 Well 9.6 57 ...

INJECTION AND HOLDING PIPETTES The glass capillary tubing used should be thin walled borosilicate glass without a fibre. e.g. Clark Electromedical Instruments GC 100T - 15 (= 15 cm) GC 100T - 10 (= 10 ...

INJECTION AND HOLDING PIPETTES The glass capillary tubing used should be thin walled borosilicate glass without a fibre. e.g. Clark Electromedical Instruments GC 100T - 15 (= 15 cm) GC 100T - 10 (= 10 ...

Need 1.5-2 x 107 cells from a 2 day culture.1. Cells are harvested as normal washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold PBS.2. Take x2 Biorad curvettesA. 0.9 ml (107 cells ...

M16 Medium (Protocol obtained from Karen Austen-Reed from SS Tan Laboratory Anatomy Department)For oocyte maturation and routine culture of mouse embryos M16 culture medium is used. This medium is una ...

MEDIA AND SOLUTIONS REQUIRED FOR ROUTINE ES CELL CULTURE Routine Culturing of ES Cells ISOLATION OF PRIMARY MOUSE EMBRYO FIBROBLASTS MITOMYCIN C TREATMENT OF PMEFs GAMMA IRRADIATION OF PMEF'S PMEF FEE ...

Generating stable cell lines in HEK293Prior to transfection it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease the likelihood of the vector integrating into the ...

Flow Cytometry Analysis (Springer Lab Harvard University) Flow cytometry employs instrumentation that scans single cells flowing past excitation sources in a liquid medium. The technology can provide ...

Introduction A. CossarizzaWorkshop SponsorsMethods in analysis of apoptosis and cell necrosis.Z. Darzynkiewicz Common methods for measuring apoptosis cell death.I. Nicoletti et al.Multiparametri ...

Collect blood (75 microliters) into 1ml PBS containing 5 microM EDTA (10 microliters of 0.5 M stock) and mix immediately to prevent clotting. Keep tubes on ice. Remove RBCs from samples. This can be a ...