MethanolFixationforImmunofluorescence

Methanol fixation works by precipitating proteins, and as such it is a quick method (2-5)minutes is enough time for most antibodies/proteins). Diffuse proteins may be lost,however, in which case PFA fixation may be a better choice. We recommend that you try both types of fixation for your particular antibody/fusion protein when working out your initial immunofluorescence conditions.MeS Buffer100 mM MeS, pH 6.91 mM EG......阅读全文

Methanol-Fixation-for-Immunofluorescence

Methanol fixation works by precipitating proteins, and as such it is a quick method (2-5)minutes is enough time for most antibodies/proteins). Diffuse

Fixation-of-Embryos

MEMFA Fix10xMEMFA Salts1 part 10x MEMFA salts1 M MOPS1 part 37% formaldehyde20mM EGTA8 parts water10mM MgSO410x salts can be autoclaved and stored. Tu

Fluorescence-Procedures-fortheActin-andTubulin-Cytoskeleton-in-Fixed-Cells2

Formaldehyde FixationFix in 4% formaldehyde (16% stock EM grade) in CBS for 20 minutesRinse in TBSPermeabilize as for methanol fixationProcede as for

Fluorescence-Procedures-forthe-ActinandTubulin-Cytoskeleton-inFixed-Cells2

Actin CytoskeletonMethanol fixationFix in -20oC methanol for 1-2.5 minutesRinse in TBSPermeabilize in TBS-0.5% TX for 10 minutesRinse in TBS-0.1% TX (

Generic-Fixation-for-Electron-Microscopy

Generic Fixation for Electron MicroscopyThe best way to fix a sample for electron microscopy is to follow a procedure developed and proven by others.

FIXATION-OF-PLANT-METAPHASE-CHROMOSOMES

Reagents (a) Metaphase arresting agents: Choose one of the following (Note 2) and shake vigorously to aerate before putting in living plant materi

Immunofluorescence-Labeling-of-Cells

实验概要Antibodies are an  important tool for demonstrating both the presence and the subcellular  localization of an antigen. Cell staining is a very ver

Tetrahymena-Fixation-for-Transmission-Electron-Microscopy

Tetrahymena Fixation for Transmission Electron MicroscopyPellet Tetrahymena cells in a clinical centrifuge.OPTIONAL: Suspend cells in HNMK (50 mM HEPE

Chlamydomonas-Fixation-for-Transmission-Electron-Microscopy

Chlamydomonas Fixation for Transmission Electron MicroscopySolutions:Chlamydomonas culture medium + 2% glutaraldehyde (5 ml medium + 0.9 ml 25% glutar

Fixation-and-Embedding-of-Microtubules-for-Electron-Microscopy

(This procedure can also be used for virtually any material that must be pelleted prior to fixation and thin sectioning)Primary fix:2% glutaraldehyde

Fixation-of-Cells-Cultured-in-Transwell-Dishes

Fixation of Cells Cultured in Transwell DishesTranswell culture dishes are commonly used to culture cells so that the top and bottom of the cells can

固氮作用(nitrogen-fixation)

分子态氮被还原成氨和其他含氮化合物的过程。自然界氮(N2 )的固定有两种方式:一种是非生物固氮,即通过闪电、高温放电等固氮,这样形成的氮化物很少;二是生物固氮,即分子态氮在生物体内还原为氨的过程。大气中90%以上的分子态氮都是通过固氮微生物的作用被还原为氨的。生物固氮是固氮微生物的一种特殊的生理功

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

Measurement-of-Carbon-Fixation-Rates-in-Leaf-Samples

Generation of a Light Curve To address the hypothesis concerning photosynthetic efficiency it is necessary to expose sun and shade leaves to a ran

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

Detection-of-Intracellular-Antigens-by-Flow-Cytometry

实验概要Fix and Perm  reagents are designed for use with all commercially available flow  cytometers. Alignment and compensation should be performed accor

Fluorescence-Procedures-forthe-Actin-andTubulin-Cytoskeleton-in-Fixed-Cells

Fluorescence Procedures for the Actin and Tubulin Cytoskeleton in Fixed CellsActin: Louise CramerTubulin: Arshad DesaiGeneral StrategyWe typically wor

Fluorescence-Procedures-forthe-ActinandTubulin-Cytoskeleton-inFixed-Cells1

General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,

Staining-Procedure-for-Flow-Cytometric-Detection-of-Human-Cyclins

Staining Procedure for Flow Cytometric Detection of Human CyclinsThis is a standard protocol used at Pharmingen for Quality Control testing of the ant

ICBR-Flow-Cytometry-Core-Laboratory-Paraformaldehyde-Fixation-of-Cells

BackgroundThis fixation method is good for cells labelled by fluorochrome-conjugated antibodies to membrane antigens. It will stabilize the light scat

Immunofluroescence-Technique

Fix cells in 2% formaldehyde in PBS/pH 7.4 for 15 min. at 20oC. 2% formaldehyde is made up fresh prior to use by dissolving the appropriate amount of

补体结合反应(complement-fixation-test)技术概况

可溶性抗原,如蛋白质、多糖、类脂质和病毒等,与相应抗体结合后,抗原抗体复合物可以结合补体,但这一反应肉眼不能察觉,如再加入红细胞和溶血素,即可根据是否出现溶血反应来判定反应系统中是否存在相应的抗原或抗体。这个反应就是补体结合反应。 补体结合反应是一种古老的血清学技术,Bordet和Gengou在19

Preparation-and-Staining-of-Paraffin-Sections

I. Fixation and Processing of Tissue for Paraffin SectionsA. Fixation of Tissues in 10% Neutral Buffered FormalinSacrifice animal by prescribed and ap

FISH-protocols-for-Drosophila2

 3. Methods3.1 RNA Probe Preparation1.   Different strategies can be used to prepare template DNA for synthesizing antisense RNA probes by in vitro tr

Mapping-Protein-Distributions-on-Polytene-Chromosomes-by-Immunostaining2

12. Hold the salivary glands at the common duct with tweezers.Transfer the glands to a drop of fixing solution on a siliconizedcoverslip. 13. Incubate

Double-immunofluore...

实验概要We provide a protocol for immunofluoresent double staining incubating the antibodies together.In order to be able to examine the co-distributi

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

Immunocytochemistry...

实验概要The method provides a guideline procedure for staining of cell cultures using immunofluoresence.实验步骤1. General procedure    1) Coat coverslips wit

MN-in-Human-Lymphocytes-(method-description)

MN in Human Lymphocytes (method description)Lymphocyte isolation    Lymphocytes were isolated using Ficoll-Paque density gradients. Blood was diluted

关于免疫诊断方法—补体结合试验(complement-fixation-test,CFT)的介绍

  补体结合试验(complement fixation test,CFT)该试验是在补体参与下,以绵羊红细胞和溶血素作为指示系统,来检测未知的抗原或抗体的血清学试验。有五种成分参与,分为指示系统和待检系统(已知抗原和未知抗体或已知抗体和未知抗原)。补体用新鲜豚鼠血清。方法是将已知的抗原或抗体与未