LabeledSphingomyelinSynthesis
Synthesis reaction:1) Mix purified DMSM with cyclohexylamine and 14CH3I at a ratio of 1/1.1/1.3 in 5 ml of methanol.2) Allow reaction to proceed at room temperature for atleast 18 hours in the dark.3) Check completion of reaction by running a few microliters of the reaction mix on TLC (solvent system = chloroform/methanol/ammonium hydroxide @ 60:35:8).4) Once reaction is complete, dry the mixture as much as possible ......阅读全文
Labeled－Sphingomyelin－Synthesis
Synthesis reaction:1) Mix purified DMSM with cyclohexylamine and 14CH3I at a ratio of 1/1.1/1.3 in 5 ml of methanol.2) Allow reaction to proceed at ro
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
Purification－of－Demethylated－Sphingomyelin
I. Lower, chloroform phase:1) Dry on rotovapor system with house vacuum lines. It is not necessary to dry sample completely, but sufficiently to yield
Starch－Synthesis
The primary carbohydrate used for energy storage in plants is starch, the equivalent of glycogen in animals. Both starch and glycogen are polymers of
Synthesis－of－plasmalogens
Plasmalogens are a class of glycerophospholipids that contain one vinyl-ester linked long chain alcohol group in place of one ester-linked fatty acid
Sucrose－Synthesis
The prodution of sucrose takes place in plants, to store energy produced during photosynthesis. The extraction of sucrose from plants like sugar cane
Lactose－Synthesis
The synthesis of glycosidic bonds between monosaccharides requires energy input, energy that is provided by joining sugar monomers to a nucleotide suc
Column－Purification－of－Demethylated－Sphingomyelin
Packing column:1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform.2) Place a small portion of glass wool at the base of the colu
Library－cDNA－Synthesis
Library cDNA Synthesis1° cDNA SynthesisN.B: During 1° cDNA synthesis, all steps should be carried while wearing gloves and all solutions should either
Synthesis－of－Cardiolipin－－phosphatidylinositol
Phosphoglycerides are lipids that contain glycerol with a phosphate ester on one end and two fatty acid ester side chains. Phosphoglycerides such as p
Protocol－for－dsRNA－Synthesis
实验概要 We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
SREBP－control－of－lipid－synthesis
Sterol-regulatory element binding proteins (SREBPs) play a key role in transcriptional regulation of cholesterol metabolism in response to cholesterol
De－novo－synthesis－of－pyrimidine－nucleotides
NON-Clickable Image The pyrimidine ribonucleotides, those found in RNA, are uracil and cytosine, each containing the sugar ribose and a nitrogenous ba
Universal－RiboClone®－cDNA－Synthesis－System
Universal RiboClone® cDNA Synthesis SystemThe Universal RiboClone® cDNA Synthesis System contains the reagents required for the synthesis of double-st
Culture－of－Endometrial－Explants－to－Monitor－Synthesis
Reprinted from:Hansen, P.J and Betts, J.G. (1992) Culture of endometrial explants and peri-implantation conceptuses to monitor synthesis and secretion
Preparation－of－nucleic－acid－probes
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
Cadmium－induces－DNA－synthesis－and－proliferation－in－macrophages
Exposure to divalent cadmium ions (Cd2+) is a known cancer risk factor, but the molecular mechanisms responsible for the inappropriate induction of ce
DNA合成（DNA－synthesis－）技术介绍
蛋白质概念提出：DNA多肽合成（DNA synthesis ）：按照预定核苷酸的顺序，将脱氧核苷酸逐个进行人工连接合成DNA链的方法。目前多是采用固相合成法，即是在多聚体支持物上从3′端延伸核苷酸，可自动化操作。)目前，oligo DNA合成一般都采用固相亚磷酰胺三酯法合成DNA片段，此方法具有高效
cDNA
· cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on
RNA标记
RNA labeling (Amersham Pharmacia)For the generation of radiolabelled, single stranded RNA for use as hybridization probes 32P-pCp 3' End-labeling
DNA标记
DNA标记（主要内容如下） DNA Labeling by Nick Translation Random Primed Labeling End-Labeling Purification of Labeled DNA Non-isotopic Labeling OthersDNA L
Sphingomyelin－Quantitation－Postcholine－Labeling－of－HL60－Cells
Lipid Extraction1) Following the appropriate time of treatment, transfer 4.5 ml into each of two duplicate glass pyrex tubes and maintain on ice.2) Sp
细胞遗传学——原位杂交(ISH)
In Situ Hybridization· In Situ Hybridization (jsmith1@po-box.mcgill.ca)In situ hybridization, as the name suggests, is a method of localizing,
Gelelongation－assay－for－type－II－fatty－acid－synthesis
Gel-elongation assay for type II fatty acid synthesisSrinivas KodaliAndrew GalgociSheo Singh Dr.Jun Wang Dr., jun_wang2@merck.com, Merck Research Labo
Synthesis－and－Probing－of－Membranebound－Peptide－Arrays2
Take the appropriate set of Fmoc-amino acid stock aliquots for cycle 1 from the freezer, bring to room temperature, and activate by adding DIC (4 µl p
Synthesis－and－Probing－of－Membranebound－Peptide－Arrays1
Synthesis and Probing of Membrane-bound Peptide ArraysRonald FrankDepartment of Chemical Biology, GBF (German Research Center for Biotechnology), 3812
Synthesis－of－Recombinant－Human－Cytokine－GMCSF－in－the－－Seeds－of－Transge...
We are interested in studying plant systems as vehicles for the production of recombinant proteins of clinical relevance. There are a number of po
Guide－to－Cell－Proliferation－and－Apoptosis－Methods
Chapter 1: Cell Death - Apoptosis and Necrosis1.1Introduction21.1.1Terminology of cell death21.1.2Differences between necrosis and apoptosis31.1.3Apop
DNA测序
DNA测序（主要内容如下）· Sequencing Gel Preparation· Preparation of Templates · DNA Sequencing by the Dideoxy Method· DNA Sequen
运用肽库筛选磷酸化激酶motif方法
Screening Kinase Phosphorylation Motifs Using Peptide LibrariesIsaac A. Manke and Michael B. YaffeCenter for Cancer Research, Massachusetts Institute