TandBcellIsolation

Reagents Heparin - 1000 U/ml Ficoll-Hypaque PBS RPMI-1640 supplemented with 10 mM glutamine and 15% FBS AET (0.14M) Dissolve 1.967 g AET in 35 ml di-H2O. Adjust to pH 8.0 with 1.0N NaOH. Bring volume to 50 ml with di-H2O.Store at 2-8oC. Check pH every 2 weeks. AET-treated SRBC Wash SRBC 4 times with PBS Add 4 volumes AET to 1 volume packed SRBC in a 15 m conical tube......阅读全文

T-and-B-cell-Isolation

Reagents Heparin - 1000 U/ml Ficoll-Hypaque PBS RPMI-1640 supplemented with 10 mM glutamine and 15% FBS AET (0.14M) Dissolve 1.967 g AET in 35 ml di-H

细胞分离技术(Cell-Isolation)

一、离心技术离心是研究如细胞核、线粒体、高尔基体、溶酶体和微体,以及各种大分子基本手段。一般认为,转速为10~25Kr/min的离心机称为高速离心机;转速超过25Kr/min,离心力大于89Kg者称为超速离心机。目前超速离心机的最高转速可达100Kr/min,离心力超过500Kg。(一)、差速离心(

Human-thyroid-endothelial-cell-isolation

1. Human thyroid glands were obtained at surgery from multinodular goiters.2. The tissue was incubated in 0.1% type II collagenase (Worthington) overn

Human-B-cell-isolation-and-culture

B cell isolation1. Donor blood was obtained with informed consent under guidelines issued by the Medical Ethics Committee. 2. Mononuclear cell fractio

Follicular-thyroid-cell-isolation-and-culture

Thyroid epithelial cells (also called follicular cells or principal cells) are cells of thyroid gland that are responsible for the production and

Human-B-cell-isolation-and-culture

实验概要This protocol provides a general protocol for human B cell isolation and culture.实验步骤B cell isolation1. Donor blood was obtained with informed con

Primary-brain-cell-isolation-and-culture

1. Cerebella were removed from 7-day-old mice and passed through Nitex nylon netting (80 μm pore size) into primary cell system containing 20% (v/

Isolation-of-dermal-mast-cell-(DMC)

1. Human DMC were isolated from skin by enzymatic digestion.2. Dermal tissue was obtained from patients with skin after informed consent was given.3. 

Isolation-and-culture-of-smooth-muscle-cell

1.Male Wistar rats of about 250g body wt were anesthetized with ether and were decapitated.  2.Kidneys and thoracic aorta were removed, rinsed with

Isolation-of-cell-nuclei-for-the-application-in-the-cellfree-system

Characteristics of the procedurePreparation of isolated nuclei - procedurePreparation of radioactive labeled nucleiMaterial Characteristics of the pro

Isolation-of-retinal-pigment-epithelial-cell(二)

15. This medium is replaced after 1 day with 5% serum-containing RPE medium, and subsequent changes were made every 2 to 3 days.16. After 3 to 4 weeks

Isolation-of-retinal-pigment-epithelial-cell(一)

The pigmented layer of retina or retinal pigment epithelium (RPE) is the pigmented cell layer just outside the neurosensory retina that nourishes

Mouse-B-cell-isolation-with-magnetic-beads

实验概要Isolate B cells from total mouse cells实验步骤 Praparation:1.       Enough MACS; put RPMI into water bath.2.       Put some MACS in 15 ml tube or 1.5

RNA-Isolation-From-Animal-tissue-or-cell-culture

实验概要This method is  designed for most animal tissues and culture cells. For RNA isolation  from fibrous tissue, follow the specialized protocol on pag

Isolation-and-Culture-of-Human-Brain-Tumor-Stem-Cell

The isolation, culture, identification, and purification of stem cells from primary human brain tumors of different phenotypes have marked capacit

Isolation-of-Cell-Wall-Proteins-from-Medicago-sativa-Stems

Plant cell walls are highly dynamic and chemically active components of plant cells. Cell walls consist primarily of polysaccharides, with protein

Mitochondrial-DNA-Isolation-from-Somatic-Embryogenic-Cell-Cultures-of-Larix

Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) and Radetzky (1990). Cell cultures at four days

Isolation-of-mouse-embryos

1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome

mice-islet-isolation

1. Islets of Langerhans were isolated from 5- to 7-week-old nonobese diabetic (NOD) mice. 2. Which involves cannulation of the common bile duct and

Yeast-Nuclei-Isolation

This method gives yeast nuclei which look nearly purified microscopically. Nuclei isolated in this way do not give active transcription extracts when

Isolation-of-murine-splenocytes

OverviewIn order to study spleen cells (e.g. lymphocytes, granulocytes, other immune cells), it helps to make single-cell suspensions so that the cell

Isolation-of-liver-lymphocyte

Isolation of liver lymphocyte   Several lymphocyte subpopulations reside in the normal adult human liver. Liver lymphocytes mainly include a large n

Isolation-of-papillary-cells

Isolation of renal papillary cells1. For isolation of papillary cells, kidneys were harvested and kept in HBSS containing 15 mM HEPES, penicillin/

Isolation-of-Zebrafish-embryos

Zebrafish will mate and deposit fertilized eggs on the bottom of the tank at 'dawn'. They can be accustomed to lay at any convenient time by k

Fungal-DNA-Isolation

Fungal DNA IsolationSaghai-Maroof MA, Soliman KM, Jorgensen RA, & Allard RW (1984) PNAS 81:8014-8018DNA successfully isolated from fungal species of C

Arabidopsis-gDNA-isolation

This is a simple and fast protocol for the extraction of genomic DNA from Arabidopsis thaliana that works fine in PCR for simple amplicons. We only us

Cosmid-DNA-Isolation

实验概要Isolation of high yields of highly pure cosmid DNA using PureLink™ HiPure Plasmid Purification Kits.实验原理The  PureLink™ HiPure Plasmid Purification

Isolation-of-colonic-epithelium

The method we use is based on work of Dr. Hazel Cheng, at the University of Toronto and works for both colon and the small intestine.First we excise t

Chromosomal-DNA-Isolation

Chromosomal DNA IsolationMETHOD:Grow cells in 2-5 ml broth to late log phase.Pellet 1-2 ml cells in microfuge.Resuspend cells in 400 µl TES (50 mM Tri

Isolation-Of-PCR-Products

实验概要Rapid and efficient purification of PCR products from salts, primers, dNTPs, and other non-nucleic acid reagents.实验原理The ChargeSwitch® TechnologyT