细胞功能测定

The 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 medium or PBS add 2 ul ...

CAM ASSAYShell-less embryo cultureFertilized white leghorn chicken eggs (SPAFAS Inc. Norwich CT) were received at day 0 andincubated for 3 days at 37°C with constant humidity. On day 3 eggs were rinse ...

PREPARING CELLSBring up HUVEC and fibroblasts in M199/10% FBS/Pen-Strep (1:100) 1-2 days before beading. Switch medium to EGM-2 (Clonetics) the day before beading for HUVEC and the day before embeddin ...

ProceduresCover a 48-well plate with Matrigel (100μl/well) of and incubate for 30 min at 37°C 5% CO2. Sacrifice the1-2 month old mice/rats (WT/mutant) and excise thoracic aorta. Transfer the aorta in ...

8 eggs per day day 7- day 13 cut CAM wash in precooled PBS in 10 ml WASH 1 (PBS 5 mM EDTA COMPLETE) on ice cut in pieces in petri dish on ice centrifuge 4 min at 1000 rpm (table top centifuge Falcon 1 ...

Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell® 12mm Diameter 12 μm Pore Size.)Procedures:1) Coat the underside of filter inserts wi ...

1) Female Tiel-PR Tg mice and wildtype control litter-mates aged 6-9 wks wereovariectimized and allowed to recover for 7-10 days.Making estrogen and progesterone stock solutions:For estrogen (17 beta- ...

(Written for 35 mm dishes ? if smaller dishes are used volumes of matrigel must be adjusted to correct for the tendency of matrigel to adhere to the sides of the dish.)Pre-coat 35 mm dish with 200 t ...

This procedure allows detection of specific cell adhesion to glycolipids resolved on TLC plates. The steps include: (1) HPTLC of glycoconjugates (2) coating the chromatogram with polymer (3) preblocki ...

Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Laminin-1 10-12 μg/ml or FN 20 μg/ml4) 96-well-plate5) C ...

DescriptionThis assay is used for the aggregation property of cancer cells. It is a very critical parameter for measurement of cell metastasis. Factors that critically affect tumor cell metastasis are ...

DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these acquisition of a motile phenotype is one requirement for a cell ...

Many different molecules have been described to promote cell adhesion including several cell surface carbohydrate-binding proteins. Measuring cell adhesion in the convenient 96-well microtiter plate f ...

THE CELL 1. Morphological aspects of apoptosis Walter Malorni Stefano Fais & Carla Fiorentini 2. Cell cycle Miriam Capri & Daniela Barbieri THE NUCLEUS 3. Fluorescence microscopy analysis of nuclear a ...

DescriptionThis is a simple assay that can be used in any cell culture lab setup to test the effect of different compounds on endothelial cell migration. ProcedureGrow endothelial cells in complete me ...

ASTRACT We describe a rapid and quantitative flow cytometric method for determining the apoptotic or anti-apoptotic potential of a gene in various cell types. A plasmid carrying green fluorescent prot ...

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 ...

Introduction When studying induction of apoptosis via a cell surface molecule it is important to first ascertain surface expression of the molecule of interest. This can be achieved by performing flow ...

Sample preparation Wash embryos 1 x in 25% MMR Remove excess buffer Add 10 volumes "incubation buffer" i.e. 50 µl for 5 embryos Lyse the embryos by gently pipetting up and down using a large blue tip ...

IntroductionIn our lab we use the term 'cell-free system' when we talk about the examination of apoptotic activity in cytoplasmic extracts. The cytoplasmic extracts may be prepared from cells which ...