SemiQuantitativeMeasurementofProteinsbyDotBlotting
Purpose...Concentration of proteins in a crude preparations (such as culture sup) can be estimated semi-quantitatively by using "Dot Blot" method if you have both purified protein and specific antibody against it.MaterialsTBS: 20mM Tris-HCl, 150mM NaCl, pH 7.5TBS-T: 0.05% Tween 20 in TBSBSA/TBS-T: 0.1% BSA in TBS-TNitrocellulose membrane (BIO-RAD, Trans-Blot™ etc)Procedure1. Have nitrocellulose membrane (NO......阅读全文
SemiQuantitative－Measurement－of－Proteins－by－Dot－Blotting
Purpose...Concentration of proteins in a crude preparations (such as culture sup) can be estimated semi-quantitatively by using "Dot Blot" method if y
Western－杂交
Western 杂交（主要内容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
Dot－blot－protocol
实验概要A technique for detecting, analyzing, and identifying proteins, similar to the western blot technique but differing in that protein samples are
Dot－Blot－Protocol
a. Label nitrocellulose membrane (using a pencil) to identify protein elution fractions.b . Pipette 2μl from each fraction onto the membrane, allow th
Purification－of－MBP－(maLTosebinding－proteins)－Fused－Proteins
Express fusion proteins as per the GST-fused protocol up to Step 7 (Day 3). All steps in protein purification should be done at 4° C unless otherwise
Detection－and－Measurement－of－Radioactivity
Radioactive DecayIsotopes of a given element have nuclei with the same number of protons but different numbers of neutrons. Some isotopes are stable,
Detection－and－Measurement－of－Radioactivity
Liquid scintillation countingThe amount of kinetic energy in a beta particle differs from one decay to the next. However, each radioisotope has a typi
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
Measurement－with－the－Light－Microscope
Measurement with the Light MicroscopeYour microscope may be equipped with a scale (called a reticule) that is built into one eyepiece. The reticule ca
SOUTHERN－BLOTTING
Materials:Whatman 3 mm Blotting Papernitrocellulose (Schleicher & Schuell, Amersham) or nylon membrane filter (Amersham).Paper towels (preferably C-fo
Western－Blotting
1. Optional: "Renature" gel- this is thought to permit some refolding of proteins and may be important in finding epitope recognition of monoclonal an
Western－Blotting
实验概要Western Blot (AP)主要试剂1. Membrane Blocking buffer:5% Milk 0.05% of Tween 20 PBS2. antibody dilution buffer: PBS 0.05% of Tween20 1.0% Milk(or BSA
dot－blot的详细步骤
蛋白质印迹法（免疫印迹试验）即Western Blot。它是分子生物学、生物化学和免疫遗传学中常用的一种实验方法。其基本原理是通过特异性抗体对凝胶电泳处理过的细胞或生物组织样品进行着色。通过分析着色的位置和着色深度获得特定蛋白质在所分析的细胞或组织中表达情况的信息。
Immunoblotting－(Western－Blotting)
实验概要We provide a protocol for SDS-PAGE, Protein Blotting, Immuno-Detection.主要试剂1. 0.3 M TRIZMA® base (Product No. T1503), 20% methanol.2. 0.025 M TRIZ
Histone－blotting－protocol
实验概要 Western blot detection of histone proteins. 实验步骤 The following protocol refers to the western blot detection of histone proteins derived from p
Western－Blotting－Protocol
实验概要The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used to detect specific proteins in the given s
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
Synaptic－Proteins－at－the－Synaptic－Junction
The postsynaptic density (PSD) is a submembranous structure at the postsynaptic membrane mainly at the excitatory synapses. The neurotransmitter recep
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
斑点杂交（Dot－blot）法
斑点杂交（Dot blot）是将被检标本点到膜上，烘烤固定。这种方法耗时短，可做半定量分析。一张膜上可同时检测多个样品，为使点样准确方便，市售有多种多管吸印仪（Manifold），如MinifoldⅠ和Ⅱ、Bio-Dot（Bio-Rad）和Hybri-Dot,它们有许多孔，样品加到孔中，在负压下
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement
IntroductionApoptosis is a normal physiological phenomenon put forward by Kerr [1]. It plays an important role in embryonic development, maintenance o
Measurement－of－Green－Fluorescent－Protein－Expression
ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 s
一种基于DNA的通用型蛋白检测系统
摘要：基于抗体的蛋白质检测方法，主要有western blot、ELISA、点杂交以及免疫组化等，这些方法被广泛地应用于科研和诊断领域。在蛋白质的免疫检测过程中，样品蛋白首先结合与特异性一抗上，然后再用携带标签（诸如：荧光染料，放射性元素，酶等）的二抗进行检测。然而，为了避免种间内的交叉反应，必
Southern－Blotting:－DNA－Transfer
Southern Blotting: DNA Transfer1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels), 7' (large gels)2. Denaturation: Wash
Western－blotting样品准备
实验概要Preparation of lysis buffers, protease and phosphatase inhibitors, lysate from cell culture, lysate from tissues, protein concentration, samples
Histone－blotting－pr...
实验概要The method provides a procedure and tips for detecting histone proteins.The protocol refers to the western blot detection of histone proteins d
Northern－blotting操作步骤
1. 取RNA2. 将电泳槽和板，梳齿浸泡在3%H2O2中20-30分钟，并吹干3. 跑 1%琼脂糖凝胶，检测样本RNA含量4. 变性胶在桌面上利用保险膜铺出一块干净的区域，将1.95g 琼脂糖加入 110ml 的 DEPC-H2O （加热前可在三角瓶上做一个记号，在加热后把蒸发的水分补足）加热
Phosphoproteins－pr...
实验概要The following procedure provides a method of detection of phosphorylated proteins.实验步骤1. To a sample of protein solution containing 1-100 ng of
Crystallization－of－Kinesin－Family－Motor－Proteins
Motor proteins of several kinesin family groups have now been crystallized: monomeric Kinesin-1 motor domains from human, rat andNeurospora (Kull et a
Acetylation－(or－Succinylation)－of－Amino－Groups－on－Proteins
Acetylation (or Succinylation) of Amino Groups on ProteinsREFERENCE: Hanock and Benz. 1986. BBA. 860:699-707.PURPOSE: Derivitization of amino groups t