GelelongationassayfortypeIIfattyacidsynthesis
Gel-elongation assay for type II fatty acid synthesisSrinivas KodaliAndrew GalgociSheo Singh Dr.Jun Wang Dr., jun_wang2@merck.com, Merck Research LaboratoriesRelated Journal & Article InformationManuscript: 2006-01-00990CJournal: NatureArticle Title: Platensimycin is a selective FabF inhibitor with potent antibiotic propertiesIntroductionType II fatty acid synthesis (FASII) is essential to bacterial cell via......阅读全文
Acid－Phenol－Yeast－RNA－Prep
This is the preferred method for yeast RNA preparationuse Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fres
核酸提取(The－Extraction－of－Nucleic－Acid)
【实验目的】 1．掌握核酸提取的基本原理和基本方法 2．掌握检测核酸浓度及纯度的原理及方法 【实验原理】 DNA是遗传信息的载体,是最重要的生物信息分子,是分子生物学研究的主要对象,因此DNA的提取也应是分子生物学实验技术中最重要、最基本的操作。如不能有效的完成DNA提取方面的工作，那就根本谈不
Protocol－for－Trichloroacetic－Acid－(TCA)－Precipitation
IntroductionThe efficiency of nucleotide incorporation in DNA/RNA polymerization reactions (e.g. transcription, reverse transcription, and DNA replica
Omega－Oxidation
NON-Clickable Image While the main route of fatty acid metabolism is through beta-oxidation, some minor metabolic pathways such as omega oxidation als
Retos－Buffers－－Media－Book
08-15 BufferCompoundMWFinal concentration200 mlGlycine75.07 0.1 M1.5 g1 M MgCl2 10 mM2 mlKOHto pH 9H2Oto 200 mlAAcetic Acid 1 MCompoundMWFinal concent
Assessment－of－Magnaporthe－grisea－mating－type－by－spore－PCR
Isolation of DNA from filamentous fungi for PCR analysis is usually time consuming and involves use of toxic chemicals such as phenol/chloroform. In S
ACACA基因突变与药物因子介绍
乙酰辅酶a羧化酶（acc）是一个复杂的多功能酶系统。acc是一种生物素酶，催化乙酰辅酶a羧化为丙二酰辅酶a，这是脂肪酸合成的限速步骤。ACC有两种形式，α和β，由两种不同的基因编码ACCα在成脂组织中高度富集该酶在转录和翻译水平上处于长期控制之下，通过靶向丝氨酸残基的磷酸化/去磷酸化和柠檬酸盐或棕榈
ACACA基因编码功能及结构描述
乙酰辅酶a羧化酶（acc）是一个复杂的多功能酶系统。acc是一种生物素酶，催化乙酰辅酶a羧化为丙二酰辅酶a，这是脂肪酸合成的限速步骤。ACC有两种形式，α和β，由两种不同的基因编码ACCα在成脂组织中高度富集该酶在转录和翻译水平上处于长期控制之下，通过靶向丝氨酸残基的磷酸化/去磷酸化和柠檬酸盐或棕榈
ChREBP－regulation－by－carbohydrates－and－cAMP
Although insulin and glucagon play important roles in regulating the response of cells to nutrients, cells also respond to carbohydrates through trans
LOWRY－PROTEIN－ASSAY
The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-
Glucosamine－Rapid－Assay
Glucosamine Rapid AssayMRTHOD:Place sample (containing 0.5 - 10 µg GlcN) in a Pyrex screw capped tube.Add HCl to a final concentration of 2N and a fin
Actin－Capture－Assay
David AmbergDialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 .Mix 5ug actin into 50ul total volume binding buffer.Mix 5ug GST-fusio
Assay－for－the－Micrococcal－Nuclease
Method: Essentially that described by Heins et al. (1966) based upon the release of acid soluble oligonucleotides following nuclease digestion of DNA.
Needle－Assay－for－Chemotaxis
Devreotes Lab, John Hopkins Medical Institutions http://www.hopkinsmedicine.org/cellbio/devreotes/needle.htmEquipment and chemicalsZeiss inverted micr
Noble－Agar－Assay
DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble
Bradford－protein－assay
Bradford protein assayConsiderations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
Tube－formation－assay
DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays, suc
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
Pheromone－Halo－Assay
-Use sterile technique and sterile solutions throughout this method.-1. Grow a starter culture at 30 C with shaking (250 rpm) until it reaches saturat
Leaf－GUS－Assay
一、实验试剂 GUS Buffer (500 ml) 2.0478 g Na2HPO4 1.2688 g NaH2PO4 (=50 mM NaPi pH7.0) 10 ml 0.5 M EDTA (=10 mM) 0.5 g Triton X-100 0.5 g N-L
HISTONE－KINASE－ASSAY
PROTOCOLTo 1.5 mL eppendorf tubes add:200 µg of protein extract (see Western blot protocol for protein sample preps)q.s. to 300 µL with RIPA (with pro
Cell－Viability－Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
cell－proliferation－assay
cell proliferation assaybefore start:thaw cells from liquid nitrogen, grow in 75cc flask (T75) in Fischer's medium MM (maintenance medium) until c
In－vitro－Sphingomyelinase－Assay
Reagents:Lysis buffer25 mM Tris-HCl, pH 7.45 mM EDTA1 mM ATP20 µg/ml CLAP1 mM PMSFBuffer A10 mM MgCl20.2 M Tris-HCl, pH 7.40.2 % Triton X-100Buffer B0
ELISA－Inhibition－Assay
ELISA Inhibition AssaySensitize a 96-well microtiter plate with purified antigen.Prepare a solution of the purified antigen of interest in phosphate b
Protein－Assay－(Spectrophotometer)
Protein Assay (Spectrophotometer)Use BSA (bovine serum albumin) 1mg/ml stock solution (1ml Eppendorf tubes) for standard curve.Place 0, 2, 5, 10, 15,
Wound－healing－assay
The wound healing assay allows the researcher to study cell migration and cell interactions. In some cases also single cell migration can be analyzed.