ProtocolforintracytoplasmicstainingofcytokinesforFACSanalysis
DescriptionProtocol for intracytoplasmic staining of cytokines for FACS analysis Procedure1) Prepare spleen, lymph node or T cell clone cells as single cell suspension, using erythrocyte lysis (water) or Lymphopaque as appropriate. 2) Stimulate cells for 2 hours in vitro with 50 ng/ml PMA (Sigma, P8139) + 500 ng/ml ionomicin (Sigma I0634) in phenol red free RPMI 1640 medium + 10% fetal calf serum at 37 deg ......阅读全文
组织学——显微解剖
Laser Capture Microdissection (LCM)Introduction to LCM (BJMU) Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections (NIH Laser Capture M
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
Combined－Flow－Cytometric－Measurement－of－Two－CellSurface－Antigens2
DNA and RNA Staining6. Stain cells with 7-AAD: i. Resuspend the cells from Step 5 in 0.5 mL of NASS containing 10 µg/mL of 7-AAD. Incubatefor 20 min a
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
侧群干细胞（sp细胞）的分离与分选
准备工作： DF12 加入0.1%BSA，100ml,取10ml冰浴，40ml放入孵箱；其余的放入4度； DNase I 100*; PI：50ug/ml; Hoechst33342: 1000ug/ml（all from Sigma） 计数板，冰盒；各种离心管，无菌流式管（BD），400滤网（BD
Intracellular－Immunofluorescent－Staining－for－Flow－Cytometry2
Table 2: Human Cytokines: Intracellular Staining Quick GuideHuman Cytokines: Intracellular Staining Quick GuideHuman CytokineCell SourceActivationIncu
Vybrant®－DyeCycle™－Violet－Stain
实验概要Live cell studies of cellular DNA content and cell cycle distribution are useful to detect variations of growth patterns due to a variety of phy
Detection－of－Intracellular－Antigens－by－Flow－Cytometry
实验概要Fix and Perm reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed accor
蛋白质检测
· Protein detection (Aberdeen's Lab)The method used to locate the proteins following 2D-PAGE depends on the nature of the original sample.
LCM－PROTOCOLS
Slide SectioningParaffin blocks- For DNA analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap™, rinsed thoro
Flow－Cytometric－Analysis－Of－Bcl－Family－members
DescriptionCell Fixation, staining and flow cytometric analysis ProcedureCells (106) were washed twice in FACS buffer (phosphate buffered saline PBS p
慢病毒转染肝细胞方法
Lentivirus Transduction of Hematopoietic CellsMing-Jie Li and John J. RossiDivision of Molecular Biology, Beckman Research Institute of the City of Ho
Cell－Sorting－By－FACS
Currently, the Moflo instrument (Dakocytomation) is used to sort cells at the AECOM FACS facility.Sorting is performed by the person in charge at the
Vybrant®－DyeCycle™－Green－and－Orange－Stains
实验概要Live cell studies of cellular DNA content and cell cycle distribution are useful to detect variations of growth patterns due to a variety of ph
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
DAPI－Counterstaining－Protocols
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding
Western－杂交
Western 杂交（主要内容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
FACS－Procedures－for－Apoptosis－Detection
Materials:Hoechst 33258 (Sigma B-2883).stock: 10 mg/ml in dH20 (40)working dilution: 500µg/ml (50µl stock + 950µl PBS).7-Amino-actinomycin (Sigma A-94
免疫组织化学
· Double Peroxidase (HRP) Immunohistochemical Labeling of Trypsin-Sensitive Antigens (KPL)· Immunohistochemistry (Tyner lab)This is a
Glycosphingolipid－analysis
1) Incubate cells with 1 µCi/ml of 3H-galactose for 72 hours.---> If treatment is for an extended period of time: treat in serum free media containing
Lipid－analysis
Thin layer chromatography is based on the separation of a mixture of compounds as it migrates with the help of a suitable solvent through a thin layer
Wholemount－staining－of－embryos
Fix embryos in formalin or MEMFA for one hour at room temperature with mixing. Rinse with TBS, replace with methanol, store at -20oC.Rehydrate by slow
Staining－Methods－for－cell
death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidiu
Protein－Staining－Procedures
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
Alkaline－phosphatase－staining
4.5.1.1 General informationEndothelial cells possess an endogenous alkaline phosphatase (AP) activity. The enzymatic activity of AP is not restricted
Fluorescent－Staining－of－Cells
1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large coverslip.2. PB
Isolation－of－human－multipotent－mesenchymal－stem－cells－from－second
Isolation of human multipotent mesenchymal stem cells from second‐trimester amniotic fluid Culture of MSC from amniotic fluid1. Twenty amniotic flui
ex－vivo－expanded－endothelial－progenitor－cells
Cell Culture. 1. Total hPBMCs were isolated from blood of human volunteers by density gradient centrifugation. 2. Cells were plated on culture dishes
DAPI－Nucleic－Acid－Stain
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding
Enrichment－of－PBMCs－with－monocytes－(The－Science－Advisory－Board)
DescriptionThis protocol is used in our lab to reduce the costs of the cell sorting with MACS reagents. The cell suspension obtained after this protoc