MeasurementofGreenFluorescentProteinExpression
ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 stock solution (1mg/ml) (see recipe)12 X 75 mm culture tubesVortex mixerWaterbath at 37oCMethod1. Count cells.2. Place approximately 106 cells into a 12 x 15 mm test tube and spin them down by centrifugation for 5 min at 300 x g.3. Remove supernatant by aspirat......阅读全文
Immunocytochemistry...
实验概要The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is a versatile marker for monitoring physiological processes, visualizi
叶绿素荧光技术：常量和微量营养元素胁迫测量
植物生长与各种常量和微量营养元素密切相关，所以测量各种常微量元素对植物的胁迫效应具有很重要的意义。大量的研究表明，Fv/Fm、Yield和ETR是与植物CO2固定能力密切相关的三个参数，所以在研究中倍受瞩目。但我们应该知道，植物营养元素的胁迫除非到了非常严重或者较为严重的程度才会影响到光系统II。这
线粒体荧光探针大全：TMRM，Mitotracker，JC1（1）
线粒体荧光探针信息大全 （Probes for Mitochondria）包括各种常用探针，如JC-1，JC-9，TMRM，TMRE等Mitochondria are found in eukaryotic cells, where they make up as much as 10% of th
Adiponectin－Replenishment－Ameliorates－ObesityRelated－Hypertension（二）
Methods Animal and Animal Treatment KKAy male mice were purchased from Japan CLEA (Tokyo, Japan). This strain is a cross between black KK fema
Protein－Immunolocalization－in－Maize－Tissues
The analysis of gene expression at transcript and at protein level is of outstanding importance in plant developmental biology. Proteins can be loc
Live－imaging－with－Drosophila－tissue－culture－cells1
IntroductionLive imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows follow
pEGFPC1质粒图谱及信息
pEGFP-C1Restriction Map and Multiple Cloning Site of pEGFP-C1. (Unique restriction sites are in bold.) The Xba I and Bcl I sites (*) are methylated in
pEGFPN1质粒图谱及信息
多克隆位点区Restriction Map and Multiple Cloning Site of pEGFP-N1. (Unique restriction sites are in color or bold.) The Not I site follows the EGFP stop cod
Detection－and－Measurement－of－Radioactivity
Radioactive DecayIsotopes of a given element have nuclei with the same number of protons but different numbers of neutrons. Some isotopes are stable,
Measurement－with－the－Light－Microscope
Measurement with the Light MicroscopeYour microscope may be equipped with a scale (called a reticule) that is built into one eyepiece. The reticule ca
Detection－and－Measurement－of－Radioactivity
Liquid scintillation countingThe amount of kinetic energy in a beta particle differs from one decay to the next. However, each radioisotope has a typi
Sphingomyelin－Mass－Measurement
Protocol:Bligh & Dyer extraction1) Pellet approximately 1 X 107 cells.2) Resuspend pellet in 3 ml CHCl3: CH3OH (1:2) and vortex hard.3) Add 0.8 ml H2O
荧光定量PCR试剂盒
荧光定量PCR试剂盒 荧光定量PCR又可以分为TaqMan探针和SYBR Green I荧光染料两种方法。比较而言，探针杂交技术在原理上更为严格，所得数据更为精确；荧光染料技术则成本更为低廉，实验设计更为简便。在选择实验方案时要根据实验目的和对数据精度的要求来决定。1. TaqMan探针法是高度特异
利用人工组合转录因子对人类基因组扫描
Scanning the human genome with combinatorial transcription factor librariesPublished online: 18 February 2003, doi:10.1038/nbt794March 2003 Volume 21
光法测定DNA浓度
Measurement of DNA concentration using PanVera fluorescence assaySteve HahnLast updated 9/30/98This Dye measures DNA concentration using a fluorescent
In－Vivo－Imaging－of－Far2
In vivo bio-imaging Mice were anesthetized and placed in a custom-made bed, which allowed stable and reproducible imaging of the legs. In vivo scann
Basic－Fluorescent－in－situ－Hybridization－(FISH)
实验概要Fluorescence in situ hybridization method is a kind of physical map drawing method, use fluorescent element mark probe, to detect probe and spli
简述PS（磷脂酰丝氨酸）在细胞外膜上的检测
PS从细胞膜内侧转移到外侧在细胞受到凋亡诱导后不久发生，可能作为免疫系统的识别标志。AnnexinV，一个钙依赖性的磷脂结合蛋白，能专一性的结合暴露在膜外侧的PS，再通过简单的显色或发光系统进行检测。由于这是一种凋亡早期的活细胞检测（悬浮细胞和贴壁细胞都适用），可与DNA染料或别的晚期检测方法相
一种基于DNA的通用型蛋白检测系统
摘要：基于抗体的蛋白质检测方法，主要有western blot、ELISA、点杂交以及免疫组化等，这些方法被广泛地应用于科研和诊断领域。在蛋白质的免疫检测过程中，样品蛋白首先结合与特异性一抗上，然后再用携带标签（诸如：荧光染料，放射性元素，酶等）的二抗进行检测。然而，为了避免种间内的交叉反应，必
Alcian－green－stain－for－cartilage
This is an easy method for stianing cartilage that can be used with isolated or CAM-grafted limbs to study limb development or with whole embryos (8 t
荧光测DNA浓度
Steve HahnLast updated 9/30/98This Dye measures DNA concentration using a fluorescent dye which binds only double stranded DNA. The fluorescence of th
关于细胞凋亡的早期检测—－PS（磷脂酰丝氨酸）在细胞外膜上的检测介绍
PS（磷脂酰丝氨酸）在细胞外膜上的检测：PS从细胞膜内侧转移到外侧在细胞受到凋亡诱导后不久发生，可能作为免疫系统的识别标志。AnnexinV，一个钙依赖性的磷脂结合蛋白，能专一性的结合暴露在膜外侧的PS，再通过简单的显色或发光系统进行检测。由于这是一种细胞凋亡早期的活细胞检测（悬浮细胞和贴壁细胞
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants. It should
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants主要试剂 Protein
pEGFPC2质粒图谱及信息
多克隆位点区（MCS）Restriction Map and Multiple Cloning Site of pEGFP-C2. (Unique restriction sites are in color or bold.) Note that the Eag I site is not uni
A－rapid,－quantitative－and－inexpensive－method－for－detecting－apoptosis2
Figure 1.Determination of apoptosis in transiently transfected murine [beta] tumor cells. (A) The number of apoptotic [beta]HC 13T tumor cells (% apop
Peripheral－blood－“endothelial－progenitor－cells”
EPC Isolation and Characterization1. EPCs were obtained by isolating mononuclear cells using Ficoll density-gradient centrifugation of human blood buf
Degenerate－PCR,－a－short－guide.
What is degenerate PCR? Degenerate PCR is in most respect identical to ordinary PCR, but with one major difference. Instead of using specific PCR pr
荧光微球(Fluorescent－microsphere)介绍
何为荧光微球(Fluorescent microsphere)？ HG-98免疫荧光分析仪除了检测带有荧光素的试剂外，还常常用于检测带有荧光微球的试剂。何为荧光微球？荧光微球: 荧光微球通常是指形状为球形，直径在几纳米至几十微米之间，微球表面或内部负载有荧光物质，在受到一定的能量激发时能够发出荧
荧光抗体(fluorescent－antibody)的制备
一、荧光素与抗体结合 1、透析法 （1）概述：适用于蛋白质含量低、样品体积小的抗体溶液的标记。标记较均匀，非特异荧光较低。 （2）步骤：将含10mg/ml的Ig溶液10ml装入透析袋，将上述透析袋放入烧杯中，含0.1mg/ml FITC的pH9.4的碳酸盐缓冲液100ml；4℃磁力搅拌24h，取出透