荧光分析法(fluorescenceanalysis

荧光光谱基础; 蛋白质的荧光特性; 荧光分光光度计的结构和原理。吸收光谱和荧光光谱能级跃迁示意图 (一)荧光的产生 某些物质受紫外光或可见光照射激发后能发射出比激发光波长较长的荧光。此化学物质能从外界吸收并储存能量(如光能、化学能等)而进入激发态,当其从激发态再回复到基态时,过剩的能量可以电磁辐射的形式放射(即发光)。荧光发射的特点是:可产生荧光的分子或原子在接受能量后即刻引起发光;而一旦停止供能,发光(荧光)现象也随之在瞬间内消失。 可以引起发射荧光的能量种类很多,由光激发所引起的荧光称为致荧光。由化学反应所引起的称为化学荧光,由X线或阴极射线引起的分别称为X线荧光或阴极射线荧光。荧光免疫技术一般应用致荧光物质进行标记。(二)荧光效率 荧光分子不会将全部吸收的光能都转变成荧光,总或多或少地以其他形式释放。荧光效率是指荧光分子将吸收的光能转变成荧光的百分率,与发射荧光光量子的数值成正比。 荧光效率=发射荧光的光量分子数(荧光强度......阅读全文

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

Analysis-of-Heme-and-Hemoproteins

Heme is perhaps the most ubiquitous cofactor found in nature and the most functionally diverse. Hemoproteins are involved in cell respiration (cyt

Flow-Cytometry-Analysis

PurposeFlow cytometry employs instrumentation that scans single cells flowing past excitation sources in a liquid medium. The technology can provide r

Lineage-Analysis-of-Blood

Materials:Capillary tubes1.5 mL Eppendorf microfuge tubes15 mL conical centrifuge tubes96-well V-bottom plates (Corning Costar 3894, from Fisher)Flow

Biosynthesis-and-Analysis-of-Bilins

The term bilin is a collective one to describe a broad group of open chain tetrapyrroles and derives from the name “bile pigments” as the first of

Analysis-of-Oligosaccharide-Ligands

Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity ChromatographyAnalysis of Oligosaccharide Ligands by High Performance Liquid 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

Analysis-of-murine-BM

Histologic analysis of murine BM is a necessary complement to flow cytometric or in vitro analysis. Techniques to do this are well established in huma

Analysis-and-Reconstitution-of-Phycobiliproteins:-Methods-for-the-...

Analysis and Reconstitution of Phycobiliproteins: Methods for the Characterization of Bilin Attachment ReactionsPhycobiliproteins are a homologous fam

Molecular-Analysis-and-Results--DNA

Theory of CGHComparative genomic hybridization (CGH) is a fairly new molecular cytogenetic technique that allows detection of DNA sequence copy number

FACS-Analysis-of-ES-Cells

Isolate cells and dissociate to single cell suspension (can use Gibco Cell Dissociation Buffer, Accutase or Trypsin)Wash with 10% FBS/DMEM:F12For surf

Protocols-for-LCM-preparation-and-analysis

Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr

Genomic-Southern-Blot-Analysis

This chapter describes a detailed protocol for genomic Southern blot analysis which can be used to detect transgene or endogenous gene sequences i

PAM:-Prediction-Analysis-for-Microarrays

PAM: Prediction Analysis for MicroarraysClass Prediction and Survival Analysis for Genomic Expression Data Mining Features:Performs sample classificat

Microtubule-Spindowns-for-Visual-Analysis

Microtubule spindowns for visual analysis can be performed on single microtubules or microtubules nucleated from axonemes/centrosomes. Although live D

Simultaneous-analysis-of-DNA-content

Simultaneous analysis of DNA content and surface immunophenotype using gentle ethanol fixation techniques.  William Telford. Louis E. King and Pamela

SOLID-TUMOR-CULTURES-FOR-CHROMOSOME-ANALYSIS

I. Purpose:A. Samples of solid tumors or lymph nodes may be sent from patients with cancer. These samples should be processed directly and also set up

TISSUE-FIBROBLAST-CULTURES-FOR-CHROMOSOME-ANALYSIS

I. Purpose:A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspected

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

TISSUE-FIBROBLAST-CULTURES-FOR-CHROMOSOME-ANALYSIS

实验概要        Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspect

Lipoprotein-Analysis-Week-2:-Electrophoresis

Lipoprotein Analysis  Week 2: Electrophoresis IntroductionSDS polyacrylamide gel electrophoresis (SDS PAGE) will be used to assess the purification pr

Reverse-Transfection-for-Gene-Function-Analysis

This guide describes a microarray-based system for the functional analysis in mammalian cells of many genes in parallel. Mammalian cells are cultured

Lipid-analysis:EXTRACTION-OF-LIPIDS-FROM-LIPOPHORIN

Lipid analysisEXTRACTION OF LIPIDS FROM LIPOPHORIN Since lipids are hydrophobic, they are better soluble in organic solvents than in water. Because th

FIXATION-and-DNA-Staining-for-Cell-Cycle-Analysis

BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to ente

Culture-of-Peripheral-Blood-Lymphocytes-for-Chromosome-Analysis

实验概要Provide information about chromosomal abnormalities.实验原理The  blood cell karyotyping method was developed to provide information  about chromosomal

Cell-cycle-analysis-of-Escherichia-coli-cells

Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of a round o

Whole-mount-TUNEL-analysis-of-Xenopus-embryos

Fixation and pretreatmentDejelly albino embryos carefully in 2% Cystein (pH 7.8).Remove the vitellin membrane with two pairs of tweezers         (or c

Protocol-for-intracytoplasmic-staining-of-cytokines-for-FACS-analysis

DescriptionProtocol for intracytoplasmic staining of cytokines for FACS analysis Procedure1) Prepare spleen, lymph node or T cell clone cells as singl

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