Wholemountantibodystainingofzebrafishembryosformarkersofsegmentation
1. Dechorionate 26 hr embryos (pharyngula stage) carefully with two fine forceps.Transfer to fixative (1% formaldehyde in PBS). Fix for 1 hour rocking at 4oC.2. Wash with 5 ml 0.1% BSA in PBS for 10 minutes. Wash 3X with 5 ml PBS,10 minutes each.3. Incubate overnight at 4oC (cold room) in 0.5 ml primary antibody in 0.2% saponin in PBS.Primary antibodies: (A) znp-1 @ 1/2000 (primary motoneurons)(B)F6@ 1/500 (somite bo......阅读全文
Whole－mount－antibody－staining－ofzebrafish－embryos－formarkers－ofsegmentation
1. Dechorionate 26 hr embryos (pharyngula stage) carefully with two fine forceps.Transfer to fixative (1% formaldehyde in PBS). Fix for 1 hour rocking
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
Zebrafish－whole－mou...
实验概要Whole mount staining of Zebrafish embryos, now commonly used, requires extra steps to fix and permeabilize to ensure the egg membrane is perme
Chick－or－Mouse－embr...
实验概要The following procedure describes the procedure for whole mount staining of chick or mouse embryo’s. A similar procedure could be used for sta
免疫组织化学
· Double Peroxidase (HRP) Immunohistochemical Labeling of Trypsin-Sensitive Antigens (KPL)· Immunohistochemistry (Tyner lab)This is a
Whole－mount－fluores...
实验概要The method provides a protocol for whole mount fluorescent immunohistochemistry. The advantage of using fluorescence to stain whole mount sectio
Immunofluorescence－Whole－Mount
Objective:Immunohistochemistry allows visualization of antigens (usually proteins) within an embryo. Typically, a primary antibody binds specifically
IMMUNOHISTOCHEMISTRY－ON－WHOLE－MOUNT－EMBR
Dechorionate directly on apple juice/agar plates with 50-60 chlorox; embryos will float to the surface after 2-3''.Collect embryos on a Millip
Wholemount－staining－of－embryos
Fix embryos in formalin or MEMFA for one hour at room temperature with mixing. Rinse with TBS, replace with methanol, store at -20oC.Rehydrate by slow
Early－development－of－primary－motor－neurons－and－somites－in－Zebrafish－Embryos
Background:Zebrafish,or the teleost fish Danio rerio,is a rapidly developing organism that is apopular species for studying vertebrate development. Cl
Immunofluorescent－staining－of－Sea－Urchin－embryos
1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 1
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
Preparation－of－fixed－embryos－for－immunocytochemistry－and－AP－staining
1. Transfer 50 ml of embryo cultures to centrifuge tubes. Spin at 1500 rpm for 5 minutes. Check that you can see a pellet of embryos at the bottom.Qui
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,
Histochemical－staining－of－sea－urchin－embryos－for－(AP)－enzyme－activity
Histochemical staining of sea urchin embryos for alkaline phosphatase (AP) enzyme activity1. Obtain embryo samples, tube of AP substrate buffer and tu
Look－Ma,－No－Archenteron!－Sulfates－role－in－sea－urchin－early－development
ObjectiveTo observe the role sulfate plays in sea urchin gastrulation, and to replicate the findings of Karp and Solursh, that sea urchin embryos fail
E.－Immunohistochemi...
实验概要We provide a guideline procedure and tips for staining of paraffin embedded sections, including antigen retrieval, chromogenic detection and flu
发育生物学
In Vitro Production of Bovine Embryos (P.J. Hansen Lab, Dept. of Animal Sciences, University of Florida)This protocol describe procedures for in vitro
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
Cell－Surface－Immunofluorescence－Staining－Protocol
实验概要A method of identifying and enumerating specific cell types in a heterogeneous population of cells by enhancing the specific staining of desired
The－Effects－of－NiCl2on－Spicule－Formation
The Effects of NiCl2on Spicule FormationJessica Ann Billet, Franklin and Marshall, Class of 2000Background and ObjectiveSea urchins exhibit radial hol
Intracellular－Cytokine－Staining－Protocol
实验概要A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surf
Application－Note:－Qdot®－Nanocrystal－Conjugates－in－Flow－Cytometry
实验概要Researchers today are trying to maximize the information that they get out of flow cytometry experiments by looking at more parameters in a sing
Immunohistochemistry
实验概要The following procedure describes the application of peroxidase or alkaline phosphatase conjugates in the immunohistochemical labeling of forma
Embryo－Lysates－－Immunoprecipitation
Embryo lysatesTake 25 embryos and place into 1.7ml centrifuge tube.Rinse once in lysis buffer (add ~ 1ml) and remove by aspirationAdd 500 µL lysis buf
In－situ－hybridization
Note: Although it is possible to perform in situ hybridization on bleached embryos, it appears to reduced the strength of the signal. For best results
The－Effects－of－Ultraviolet－Light－on－the－Fertilization
Jill K. Flemming, Franklin & Marshall College, Class of 2001IntroductionThe objective of this project is to observe the effects of UV radiation on bot
骨组织βgal免疫染色实验技术方法
ß-gal Staining:Reagents:0.1M Phosphate Buffer (pH 7.3):0.1M Sodium Phosphate monobasic115ml0.1M sodium phosphate dibasic385mlTotal Volume500 mlFix sol
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 (
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