FlowSortingFibroblastswithGFPandP.I.
ProtocolWash cells with PBS and trypsinize to a single cell suspension.Count an aliquot on a hemocytometer. Meanwhile centrifuge the cells (e.g. 1000 RPM, 5 min.) to pellet them.Resuspend the cells to a concentration of 2 x 106 cells/sample with Flow Sort solution.Aliquot into flow sort tubes (maximum 3 mL per tube)Aliquot 1.5 mL of media into falcon tubes for collection.Place cells and collection tubes on ice a......阅读全文
Flow－Sorting－Fibroblasts－with－GFP－and－P.I.
ProtocolWash cells with PBS and trypsinize to a single cell suspension.Count an aliquot on a hemocytometer. Meanwhile centrifuge the cells (e.g. 1000
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
Virus－Infection－of－Fibroblasts
Procedure: Day One 1. Plate cells in 60 mm dishes using 5 ml DMEM containing 10% Calf Serum. The protocol has worked well for NIH3T3 with 1 x 105 cell
Virus－Infection－of－Fibroblasts
Procedure: Day One 1. Plate cells in 60 mm dishes using 5 ml DMEM containing 10% Calf Serum. The protocol has worked well for NIH3T3 with 1 x 105 cell
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
Keratocyte－isolation－(corneal－fibroblasts)
Corneal keratocytes (corneal fibroblasts) that this corneal layer is specialized fibroblasts residing in the stroma, representing about 85-90% of
Dynamic－Flow－Assay－in－a－Parallel－Plate－Flow－Chamber
Dynamic Flow Assay in a Parallel Plate Flow ChamberJohn T. Patton~GlycoTech Corporation, Rockville, Maryland 20850Flow assays allow visualization of c
GFP抗体|GFP抗体检测GFP、EGFP、YFP、EYFP、CFP抗体
检测GFP、EGFP、YFP、EYFP、CFP的GFP抗体GFP是绿色萤光蛋白（Green Fluorescent Protein）的简称，由238个氨基酸残基组成。GFP蛋白质最早是由下村脩等人在1962年在一种学名Aequorea victoria的水母中发现。其基因所产生的蛋白质，在蓝色波长范
ISOLATION－OF－PRIMARY－MOUSE－EMBRYO－FIBROBLASTS
You will need:13.5 day pregnant mouse (we use MTK NEO inbred white mice)2 sets sterile instrumentsone containing a pair of curved forceps and a pair o
Isolation－of－rat－cardiac－fibroblasts－and－cardiomyocytes
1. Hearts were removed from newborn rats (day 0), put into calcium- and bicarbonate-free HEPES-buffered Hanks’ medium, cut into pieces and digeste
Purification－of－Lin,－ckit+,－Sca1+－bone－marrow－cells－for－Culture
MATERIALSMedia:Heat inactivated FBS (56°C x 30 min)PBS + 2% heat-inactivated FBSIMDM + 20% heat-inactivated FBSViral transduction: Iscove's + 20%
Flow－Cytometry－Analysis
PurposeFlow cytometry employs instrumentation that scans single cells flowing past excitation sources in a liquid medium. The technology can provide r
Isolation－and－growth－of－pulmonary－artery－adventitial－fibroblasts
1. Adventitia from the main pulmonary artery was harvested neonatal calves. 2. Tissue was collected, carefully dissected free of blood vessels and f
什么是－Flow－Chemistry－？
flow chemistry 一般译作 流动化学。本意指在连续流动的系统中完成化学反应，不同于批次式反应。其实流动过程中完成化学转化的生产方式并不独特，早已广泛用于石化工业和 合成氨、硫酸、盐酸、硝酸等大化工领域。真正让流动化学独具魅力的是 小型化 和 智能化。流动化学不仅仅单纯指物料的流动，而是结
Hematopoietic－Stem－Cell－Targeting－with－Surface1
Hematopoietic Stem Cell Targeting with Surface-Engineered Lentiviral VectorsEls Verhoeyen and François-Loïc CossetAdapted from Gene Transfer: Delivery
GFP单抗的概述
GFP单抗，又叫GFP单克隆抗体，或维多利亚水母绿色荧光蛋白单抗，是蛋白质研究过程中非常重要的工具，尤其是在鉴定重组的带有GFP标签的蛋白质是否表达或者表达的相对丰度时有着极重要的作用。
Yeast－Cell－Cycle－by－Flow－Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4
Intracellular－Immunofluorescent－Staining－for－Flow－Cytometry
IntroductionA modification of the basic immunofluorescent staining and flow cytometric analysis protocol can be used for the simultaneous analysis of
Flow－Cytometric－Analysis－of－Cell－Cycle
Fixation1) Collect 2 X 106 cells.2) Pellet cells by spinning at 1,000 rpm, 4°C for 5 minutes.3) Resuspend cell pellet in 1 ml of cold PBS.4) Fix cells
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
Retrovirus－Production
Material:Packaging Cells, e.g. Phoenix cells (an adenovirus Ad5-transformed human embryonic kidney cell line 293T, transfected with two MoMLV packagin
iRNA干扰GFP表达实验
实验概要本文介绍了iRNA干扰GFP表达实验的原理及方法步骤。实验原理RNA沉默是发生在植物（转录后基因沉默或共抑制）、动物（RNA干扰，RNAi）和真菌（消除作用）等真核生物细胞中的的特异性和高效率的mRNA降解机制。在哺乳动物细胞中，RNAi通常用于阻断特定基因的表达从而研究基因的功能。将靶向特
iRNA干扰GFP表达实验
【原理】RNA沉默是发生在植物（转录后基因沉默或共抑制）、动物（RNA干扰，RNAi）和真菌（消除作用）等真核生物细胞中的的特异性和高效率的mRNA降解机制。在哺乳动物细胞中，RNAi通常用于阻断特定基因的表达从而研究基因的功能。将靶向特定基因的大约21碱基长短的双链siRNAs(smallinte
Culture－of－human－renal－proximal－tubule－cells－and－renal－cortical－fibroblasts
1. The method for primary culture of human renal proximal tubule cells (PTC) and renal cortical fibroblasts (CF) is described in detail.2. Renal cor
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
Flow－Cytometry－of－Fibroblast－Nuclei－for－DNA－content
MaterialsP.I. Solution: 4 mM Na3Citrate (0.118 g/100 mL)30 U/mL RNAseI (43 mg/100 mL)0.1% Triton-X100 (0.1mL/100 mL)50 µg/mL propidium iodide (5 mg/10
A－rapid,－quantitative－and－inexpensive－method－for－detecting－apoptosis
ASTRACTWe describe a rapid and quantitative flow cytometric method for determining the apoptotic or anti-apoptotic potential of a gene in various cell
活体GFP绿色荧光成像系统
系统提供动物活体绿色荧光蛋白的实时观察与成像等一系列的荧光检测。能够应用在像深度肿瘤，大动物等活体肿瘤追踪观察成像研究。 该设备是一个高灵敏度的图像成像工作系统，主要利用特定波长的激光进行激发后，通过高灵敏度的致冷CCD进行实时检测后，获得所需的各类 特性的图像，有利于进一步的分析作用 。
GFP与YFP有哪些区别
YFP和GFP其实是序列基本相同的两种蛋白，YFP就是把Thr203以Tyr取代，GFP则不发出绿色荧光，而发出较长波长的黄色荧光，也就是YFP。因此两者最大的区别则是发射波长了。我觉得这两种蛋白标记应该都没有问题，只是有几个问题应该考虑：1. 应该标记在C端，一般的核定位序列均位于蛋白N端，如果将
GFP：荧光蛋白的起源
绿色荧光蛋白(简称GFP)，是一个由约238个氨基酸组成的蛋白质，从蓝光到紫外线都能使其激发，发出绿色荧光。GFP的荧光非常稳定，在激发光照射下，其抗光漂白能力比荧光素强很多。因此GFP及其变种被广泛地用作分子标记；此外，GFP还被用作砷和一些重金属的传感器。 1962年，下村脩和约翰逊在一