ColumnPurificationofDemethylatedSphingomyelin
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 column.3) Pour gel solution into column and use a stir bar to press out any bubbles in the glass wool.4) Slowly drain the chloroform while stirring to prevent bubbles from being present in the packed gel.5) Add 20 mls more chloroform to recover the remaining portion of sili......阅读全文
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
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
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
Antibody－Purification
This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography.1. Solutions(1) Affigel Blue Prewash0.1 M acet
COLUMN－MANAGEMENT－色谱柱管理
10.1. Columns used in chromatography should be appropriate for their intended use.色谱所用柱子应适合其既定用途。10.2. Columns should be purchased from approved supp
蛋白质提取和纯化
蛋白质提取和纯化（主要内容如下）Protein Extraction Protein PurificationProtein PrecipitationColumn PreparatioinQ & A Posted in the Method ForumProtein ExtractionWhole
抗体纯化
Antibody PurificatioinPurification of IgG Using Protein A- or Protein G-Agarose (KPL) Purifying Antibodies (Perkin-Elmer)Precipitation MethodsProtein
CD2的纯化－Purification－of－CD2
Purification of CD21) Cells are pelleted (~3 - 4000 g for 10 min.) and resuspended in pre-chilled 50 mM malonate (pH 5.2-5.3) with 1mM EDTA (~40ml of
Purification－of－mAb－(IgG)
1. Materials(1) Antibody 7E3, 2L sup grown in flasks, frozen and thawed overnight.(2) BioRad Affi-Gel Protein A MAPS II Buffers cat. #1530-6160 ($161.
Protein－purification;－actin
Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
Protein－A－Purification－of－Antibody
1. Reagents(1) Affi-gel Protein-A Agarose (BioRad #153-6153)(2) MAPS II Binding Buffer (BioRad # 153-6161)(3) 0.314 g/ml diH2O(4) MAPS II Elution Buff
Column－Method－for－Lambda－Phage－DNA－Preparation
Purpose:Mini-prep method for lambda phage DNA purification from lysates.Time required:4 hours once the lysate is in handSpecial supplies required:BioR
Cosmid－DNA－Isolation
实验概要Isolation of high yields of highly pure cosmid DNA using PureLink™ HiPure Plasmid Purification Kits.实验原理The PureLink™ HiPure Plasmid Purification
Midiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
亲和层析凝胶指南
GE HealthcareBenzamidine Sepharose™ 6B is p-aminobenzamidine covalentlyattached to Sepharose 6B by the epoxy coupling method.p-Aminobenzamidine (PAB),
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
Protein－G－Purification－of－Antibodies
1. Reagent and Materials(1) Hi-Trap Protein G Column (Pharmacia Biotech #17-0404-01)(2) 20 mM Sodium Phosphate Buffer, pH 7.01.084 g NaH2PO4, anhydrou
Antigen－Affinity－Purification－of－Antibodies
实验概要To acquire purified antibodies (This method typically yields >95% pure specific antibodies ).实验原理 Cytokines are signaling proteins necessary for
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
Purification－of－GST－Fused－Proteins
Day 1Set up an overnight culture in 100 ml LMM broth or 100 ml terrific broth containing 100ul 100 mg/mlAmpDay 2Add 40-50 ml o/n culture to 1 lt terri
AntiDYKDDDDK－tag－(L5)－Affinity－Gel
实验概要Anti-DYKDDDDK tag (L5) affinity gel is a purified rat IgG2a, κ monoclonal antibody covalently attached to agarose by hydrazide linkage. It is us
Rapid－Isolation－and－Purification－of－Photosystem－I－ChlorophyllBinding...
The available procedures for isolation and purification of photosystem I (PSI) from Chlamydomonas reinhardtii are time consuming and usually require
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
DNA－Purification－from－Agarose－Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Purification－of－human－mononuclear－cells－and－neutrophils
PurposeMaterials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS4
Sodium－Azide－removal－from－antibody－solutions
实验概要Sodium azide is a preservative used for inhibiting the growth of contaminants such as bacterial or fungus in antibody solutions. However, its
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
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
微量RNA的抽提RNeasy－MinElute－Spin－Column
·为了更好地裂解，细胞数不能大于5×105，细胞过多会减低产率和纯度。 准备工作： 1.在 24 ml 96－100％乙醇 中加入 6 ml 去RNA酶的水。 2.在每 1 ml Buffer RLT 中加入10 ml β-ME (加入β-ME的Buffer RLT可在室温下放置1