Multicolour3DFISHinvertebratecells5
Author NotesAfter the fourth round of DOP amplification the probe quality is considerably reduced.Use the low stringency cycles only in case you start with genomic DNA. For all re-amplifications omit low stringency cycles and use only high stringency cycles.In case of the simultaneous labeling of several probes (chromosome specific painting probes) with the same hapten or fluorochrome allow 1µl of unlabeled amplifica......阅读全文
Isolation－and－culture－of－pancreatic－stellate－cells
Pancreatic stellate cells (PaSCs or PSCs) are myofibroblast-like cells found in the areas of the pancreas that have exocrine function. PaSCs are
Isolation－of－human－endometrial－epithelial－cells
Tissue collection1. Endometrial biopsies were collected from women undergoing gynaecological procedures for benign conditions. 2. All women reported
Purification－of－human－mononuclear－cells－and－neutrophils
PurposeMaterials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS4
Isolation－of－normal－mammary－epithelial－cells
1. A small piece of tissue from each specimen was removed and minced. 2. The tissue was digested with collagenase overnight. 3. To remove the collagen
Activation－and－Expansion－of－Human－Treg－Cells
实验概要This protocol is intended for activation and expansion of human Treg cells isolated with the Dynal® CD4 CD25 Treg Kit (Cat. no. 113.23D). The exp
Transfection－of－Mammalian－Cells－Using－Lipofectamine
Materials: LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium cont
DNA－laddering－assay－for－treated－cells
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 c
Differentiate－ES－cells－into－cardiac－myocytes
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.____________________Day 1: Trypsini
Rat－urinary－bladder－urothelial－cells
1. Bladders were excised from deeply anesthetized (urethane, 1.2 gm • kg−1, i.p.) Sprague Dawley rats (of either sex), cut open, and gently stretc
Isolation－of－human－prostatic－epithelial－cells
1. A small piece of tissue from each specimen was removed and minced. 2. The tissue was digested with collagenase overnight. 3. To remove the collagen
Isolation－and－culture－of－porcine－muller－cells
1. Porcine eyes were harvested from pentobarbital-anesthetized animals, each weighing 30 to 40 pounds, and transported to the laboratory in ice-co
Culturing－Human－Neural－Stem－Cells
实验概要Neural stem cells (NSC) are valuable resources because of their ability to differentiate into neurons and glial cells with applications in neur
Isolation,－Culture,－and－Differentiation－of－Progenitor－Cells
Isolation, Culture, and Differentiation of Progenitor Cells from the Central Nervous SystemScott R. Hutton and Larysa H. Pevny1UNC Neuroscience Center
Fixation－of－Cells－Cultured－in－Transwell－Dishes
Fixation of Cells Cultured in Transwell DishesTranswell culture dishes are commonly used to culture cells so that the top and bottom of the cells can
Dissociation－of－Cells－from－Primary－Tissue
实验概要A common method to obtain single cell suspensions from primary tissue is enzymatic disaggregation. Expose the cells to enzymes for a minimal am
CELL－CYCLE－ANALYSIS
PROPIDIUM IODIDE: The most commonly used dye for DNA content/cell cycle analysis is PROPIDIUM IODIDE (PI). It can be used to stain whole cells or isol
Segmentation－Clock
In most animal species, the anteroposterior body axis is generated by the formation of repeated structures called segments. In vertebrate segmentation
Cryopreservation－and－Recovery－of－Mature－Differentiated－Neural－Cells
实验概要Primary neuronal cultures are indispensable in the field of neurobiology and pharmacology. Many researchers favor freshly isolated neuronal cell
Culture－of－Human－Embryonic－Stem－Cells－(hESC)
All cell lines are initially grown according to the supplier's protocols but we are adapting them to one simple protocol outlined below:6-well pla
Isolation－of－human－uterine－smooth－muscle－cells
1. Approval to utilize human tissue was given by Hospital's Committee on Research Involving Human Subjects. 2. Human myometrial and leiomyoma tiss
Eccles:Protein－Lysates－from－Cells－in－Culture
Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH
HOW－TO－USE－THE－COULTER－COUNTER－TO－COUNT－CELLS
1) Turn on the counter by pulling out the on/off button. You need to do this at least 10 min before use to obtain sufficient vacuum.Usually put 0.2 ml
Production－of－Recombinant－Proteins－in－SuspensionCultured－Plant－Cells
Plants have emerged in the past decade as a suitable alternative to the current production systems for recombinant pharmaceutical proteins and, toda
Cell－cycle－analysis－of－Escherichia－coli－cells
Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of a round o
Differentiate－ES－cells－into－cystic－embryoid－bodies
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized th
Transient－Transfection－of－Cos1－Cells
Transient Transfection of Cos-1 CellsNote: This protocol has been optimized for Cos-1 cells. Successful transfection of each cell type requires optimi
Culturing－Rat－Fetal－Neural－Stem－Cells
实验概要Rat neural stem cells (NSCs) serve as a well-established model for investigating human brain development, disease processes, and treatment stra
Cellbased－ELISA－for－primary－cells
1. The 96-well micro-plates are pre-coated with extra cellular matrix.2. Primary cells are grown in the 96-well plates. Seed 100μl of 10,000-20,000
Isolation－of－human－prostatic－smooth－muscle－cells
Human prostate tissues1.Human hyperplastic prostates were obtained during surgery from four men through transurethral resection of the prostate. 2.All
Tissue－Culture－of－PtK1－cells
Please note: lately (2005 - present), we normally culture PtK1 cells in F-12 media, but they also grow in other types of media as described below.Mixi