PreparationandStainingofParaffinSections
I. Fixation and Processing of Tissue for Paraffin SectionsA. Fixation of Tissues in 10% Neutral Buffered FormalinSacrifice animal by prescribed and approved euthanasia techniques. Tissues to be fixed and processed should be cut to a size no larger than 3mm thick. Let tissues fix in 10% formalin at room temperature for 8 hours but not to exceed 24 hours. For small rodent tissue, it is recommended to fix tissues for 4-......阅读全文
Whole－mount－fluores...
实验概要The method provides a protocol for whole mount fluorescent immunohistochemistry. The advantage of using fluorescence to stain whole mount sectio
蛋白质提取与制备(Protein－Extraction－and－Preparation)2
三、蛋白质提取与制备具体操作方法1、原料的选择早年为了研究的方便，尽量寻找含某种蛋白质丰富的器官从中提取蛋白质。但至目前经常遇到的多是含量低的器官或组织且量也很小，如下丘脑、松果体、细胞膜或内膜等原材料，因而对提取要求更复杂一些。原料的选择主要依据实验目的定。从工业生产角度考虑，注意选含量高、来源丰
蛋白质提取与制备(Protein－Extraction－and－Preparation)1
蛋白质提取与制备蛋白质种类很多，性质上的差异很大，既或是同类蛋白质，因选用材料不同，使用方法差别也很大，且又处于不同的体系中，因此不可能有一个固定的程序适用各类蛋白质的分离。但多数分离工作中的关键部分基本手段还是共同的，大部分蛋白质均可溶于水、稀盐、稀酸或稀碱溶液中，少数与脂类结合的蛋白质溶于乙醇、
反向微柱的准备Preparation－of－ReversedPhase－Microcolumns
INTRODUCTIONOne versatile strategy for sample cleanup prior to MALDI-MS analysis uses microscale columns designed for direct sample elution onto the M
蛋白质提取与制备(Protein－Extraction－and－Preparation)6
PH 值：与沉淀蛋白质或酶原理相同，结晶的溶液PH 值一般选择在被结晶的蛋白质或酶的等电点附近，以利于晶体的析出。温度：除少数情况外，通常选择低温条件进行。低温条件对蛋白质和酶不仅溶解度低且不易变性。在中性盐溶液中结晶时，温度可在0℃至室温范围内选择，在有机溶剂中结晶一般要求温度较低。晶种：不易结晶
蛋白质提取与制备(Protein－Extraction－and－Preparation)4
蛋白质提取液中，除包含所需要的蛋白质（或酶）外，还含有其它蛋白质、多糖、脂类、核酸及肽类等杂质。除去的方法有：1）核酸沉淀法该法可用核酸沉淀剂和氯化锰、硫酸鱼精蛋白或链霉素等。必要时也可用脱氧核糖核酸酶除去核酸。即在粗匀浆中加入少量DNase,于4℃保温30～60min，可使DNA 降解为足够小的碎
Preparation－of－Genomic－DNA－from－Bacteria－using－Phase－Lock－GelTM
Preparation of Genomic DNA from Bacteria- using Phase Lock GelTM (Modified from Experimental Techniques in Bacterial Genetics, Jones and Bartlet, 1990
蛋白质提取与制备(Protein－Extraction－and－Preparation)3
水溶液提取：大部分蛋白质均溶于水、稀盐、稀碱或稀酸溶液中。因此蛋白质的提取一般以水为主。稀盐溶液和缓冲溶液对蛋白质稳定性好、溶度大，也是提取蛋白质的最常用溶剂。盐溶液提取：以盐溶液及缓冲液提取蛋白质进常注意下面几个因素。盐浓度等渗盐溶液尤以0.02～0.05mol/L 磷酸盐缓冲液和碳酸盐缓冲液常用
蛋白质提取与制备(Protein－Extraction－and－Preparation)5
确定沉淀蛋白质所需硫酸铵浓度的方法将少量样品冷却到0～5℃，然后搅拌加入固体硫酸铵粉末，见蛋白质产生沉淀时，离心除去沉淀，分析上清液确定所要蛋白质的浓度，如它仍在溶液中则弃去沉淀，再加更多的硫酸铵于上清液中，直到产生蛋白质沉淀时止。以所要提取的蛋白质在溶液中的浓度对硫酸铵浓度作图，得沉淀曲线，找出蛋
美国实验室wetern方法
WESTERN BLOT PROTOCOLIn a Western blot, proteins that are separated on polyacrylamide gels on the basis of size are transferred to a membrane for dete
PCR的下游应用
・ Agarose Gel Electrophoresis of PCR Products(Robert H. Cruickshank)・ Agarose Gel Electrophoresis of PCR Products(Immunology Resource)
实验室荧光显微镜的标本制作方法与正确步骤
一、 玻片及盖玻片(Slide and Coverslips) 玻片及盖玻片质量必需很好而没有荧光，玻片厚度≦1.0mm 者便可用，玻片之清洗，必需以中性清洁剂洗净，再浸于含有3％HCl 之乙醇12～24 小时，然后再贮存于纯酒精。要用时，以清洁纱布擦净或火焰烘干。用完后可浸于水中，移去封
Isolation－of－liver－infiltrating－lymphocytes
1. Hepatic T cells were isolated from fresh liver tissue collected into primary cell system.2. In the second method, tissue was diced using sterile bl
细胞周期试剂盒Cell－cycle－staining－Kit中文操作步骤
试剂盒特点：1、独创的活细胞一步法检测，收集——洗涤——染色，简单快速 2、兼容固定细胞检测，改进的固定方法，更少细胞粘连，更小CV值 3、兼容流式和荧光显微镜分析，提供详细protocol固定细胞处理：1、收集细胞悬液，细胞数为2 × 10^5 ~ 1 × 10^6。离心沉淀细胞，去上清，
Fluorescence－Mounting－Medium－(Antifade)
Materials Needed20ml glass scintillation vialSmall stir barFoilGlycerol1X PBSPipets* P-phenylenediamine ( EMD Chemicals Inc. Cat# PX0730)Carbonate-Bic
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
Analysis－of－Proteins－using－Small－Format－2D－Gel－Electrophoresis
Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei
如何正确制作荧光显微镜标本
(一) 玻片及盖玻片(Slide and Coverslips)玻片及盖玻片品质必须很好而没有荧光，玻片厚度≦1.0mm 者便可用，玻片之清洗，必须以中性清洁剂洗净，再浸于含有3％HCl 之乙醇12～ 24 小时，然后再储存于纯酒精。要用时，以清洁纱布擦净或火焰烘干。用完后可浸于水中，移去封埋物，再
碳水化合物分析
Carbohydrate Assay (Hancock Laboratory) (Accessible only by IE)This protocol is used to determine the relative amounts of LPS CHO present in a given s
寡核苷酸的相关操作
In this section, you will find techniques related to oligonucleotides, such as oligo purification by acrylamide gel, annealing two oligos to make doub
酵母遗传学技术
Genome-wide Gene Expression Analysis (Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis u
Axolotl－Microsurgery
I. Embryo Preparation1) Manually dejelly embryo of appropriate stage. Use fine foreps to break hole in hard jelly coat. Remove membrane around embryo
ELECTROPHORESIS－OF－DNA－IN－AGAROSE－GELS
ELECTROPHORESIS OF DNA IN AGAROSE GELSA). AGAROSE CONCENTRATIONS: Use 0.8% agarose (w/v) for high molecular weight DNA fragments, and 1 - 1.2% f
FFPE组织样品的DNA和RNA纯化
福尔马林固定后石蜡包埋的组织简称为FFPE样品。将组织在 4–10% 的福尔马林中迅速固定。 固定时间限制在 14–24 小时 (越长的固定时间将越容易导致DNA的片段化，对下游实验不利)。将固定组织彻底脱水 (彻底脱去福尔马林残余，因为其阻碍蛋白酶K的作用)。 纯化DNA时，使用新鲜从石蜡块上切下
Intestinal－intraepithelial－lymphocytes
Intestinal intraepithelial lymphocytes Intestinal intraepithelial lymphocytes (IELs) are mostly T cells, which are continuously associated with gut
The－Effects－of－Ultraviolet－Light－on－the－Fertilization
Jill K. Flemming, Franklin & Marshall College, Class of 2001IntroductionThe objective of this project is to observe the effects of UV radiation on bot
Dual－and－TripleCo...－（二）
主要试剂1. Blocking reagent after coating: 10% fetal calf serum (FCS) in phosphate-buffered saline (PBS), pH 7.4.2. Cells suspended in complete culture me
CellTrace™－CFSE－Cell－Proliferation－Kit
实验概要The CellTrace™ CFSE Cell Proliferation Kit provides a versatile and well-retained cell-tracing reagent in a convenient and easy-to-use form. The
Use－of－SemiThin－Cryosections－for－Light－Microscopy.
Use of Semi-Thin Cryosections for Light Microscopy.Semi-thin sections can be obtained from frozen blocks of cryoprotected biological material by secti
IHC－Protocol－for－Fr...
实验概要The method provides a IHC protocol for free floating brain sections.实验步骤1. Coronal 30-40 µm sections cut on a freezing microtome. Sections colle