细胞功能测定

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- ...

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 ...

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 ...

Flow assays allow visualization of cell adhesion under well-defined wall shear stress. The visualization of the different events of cell adhesion can be quantified by selective image acquisition and s ...

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 ...

Rapidly thaw and immediately place on ice one aliquot each of axonemes Golgi or ER membranes 45 uM tubulin rat liver cytosol and 20x energy regeneration system. Dilute the axonemes with PM Buffer to t ...

The wound healing assay allows the researcher to study cell migration and cell interactions. In some cases also single cell migration can be analyzed. This assay can be imaged using Nikon microscope 3 ...

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 ...

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 ...

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 ...

Protocol I: Triton X-100 Lysis Buffer In 96 flat-wells plate incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of effectors (105 target cells per well). After incubati ...

DESCRIPTION of the method:The DNA Fragmentation Assay allows to determine the amount of DNA that is degraded upon treatment of cells with certain agents e.g. with TNF-alpha or anti-Fas antibody (IPO-4 ...

1. Introduction Nuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonuclease-mediated cleavage of nuclear DNA. In fact ...

I. Protocol1. Harvest cellsOptional: wash plate with 37°C PBS (gently so as not to lose cells); check on microscopeafter aspirate PBS or mediaPlace plate on iceLyse with DNA ladder buffer 4°C150 ul/60 ...

Characteristics of this procedure:I found the procedure described by Gong et al. to be a convenient and successful method to detect DNA laddering in cells undergoing apoptosis. As the authors describe ...

Protocol for Paraffin Sections:Dewax paraffin sections: Incubate slides 55°C 30 min.Xylenes 2 times 2 min. each100% EtOH 2 times 2 min. each95% EtOH 2 times 2 min. each80% EtOH 2 min.75% EtOH 2 min.50 ...

1. INTRODUCTION Apoptosis was observed from invertebrates to lower and higher verterbrates and intervenes both in physiological and in pathological states such as normal cell turnover embryogenesis ...

PROTOCOL:•Deparaffinize and rehydrate slides:3 x 3´ Xylene3 x 2´ 100% ethanol1 x 2´ 95% 80% 70% ethanol (each)1 x 5´ 1x PBS•Microwave antigen retrieval:4 x 5´ in microwave (600 ml of 10mM NaCitrate pH ...

Fixation and pretreatment Dejelly albino embryos carefully in 2% Cystein (pH 7.8). Remove the vitellin membrane with two pairs of tweezers (or carefully pierce multiple times after fixation) F ...