WholemountTUNELanalysisofXenopusembryos
Fixation and pretreatmentDejelly albino embryos carefully in 2% Cystein (pH 7.8).Remove the vitellin membrane with two pairs of tweezers (or carefully pierce multiple times after fixation)Fix the embryos in MEMPFA for 1 hour at room temperature (MEMPFA: 100 mM MOPS pH 7.4, 2 mM EGTA, 1 mM MgSO4, 4% paraformaldehyde)Wash 2 ......阅读全文
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
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
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－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
Effect－of－Cyclopamine－on－Xenopus－laevis－embryos
Cyclopamine and jervine, teratogens derived from the skunk cabbage Veratrum californicum, are known to induce holoprosencephaly in chick embryos when
Cell－death－detection－in－Xenopus－embryos－by－ELISA
Sample preparationWash embryos 1 x in 25% MMRRemove excess bufferAdd 10 volumes "incubation buffer", i.e. 50 µl for 5 embryosLyse the embryos by gentl
DEVELOPMENTAL－EFFECTS－OF－LITHIUM－CHLORIDE－ON－XENOPUS－EMBRYOS
PURPOSE: The objective of this experiment is to study the effect of lithium chloride treatment on early blastula xenopus embryos.INTRODUCTION: When th
TUNEL－PROCEDURE－IN－BOVINE－EMBRYOS
Materials8% (w/v) paraformaldehyde stock solution: Dissolve 8 g of powdered paraformaldehyde in 100 ml water. Heat and stir (55-60 C – do not go high
TUNEL－PROCEDURE－IN－BOVINE－EMBRYOS
Materials8% (w/v) paraformaldehyde stock solution: Dissolve 8 g of powdered paraformaldehyde in 100 ml water. Heat and stir (55-60 C – do not go high
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
Tunel－Procedure－in－Bovine－Embryos－牛胚胎TUNEL检测凋亡
Materials8% (w/v) paraformaldehyde stock solution: Dissolve 8 g of powdered paraformaldehyde in 100 ml water. Heat and stir (55-60 C – do not go highe
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
Dorsal－Lip－Transplant－in－Amphibian－Embryos
IntroductionIn early stages of development in amphibian embryos, very few cells' fates are rigidly determined. However, previous experiments show
免疫组织化学
· Double Peroxidase (HRP) Immunohistochemical Labeling of Trypsin-Sensitive Antigens (KPL)· Immunohistochemistry (Tyner lab)This is a
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,
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
cDNA
· cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on
Apoptosis:－A－Laboratory－Manual－of－Experimental－Methods－Andrea－Cossarizza
THE CELL 1. Morphological aspects of apoptosis Walter Malorni, Stefano Fais & Carla Fiorentini 2. Cell cycle Miriam Capri & Daniela BarbieriTHE NUCLEU
SingleCell－Electroporation－in－Xenopus
Single-Cell Electroporation in XenopusXue Feng Liu and Kurt HaasINTRODUCTIONSingle-cell electroporation (SCE) is a versatile technique for delivering
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
Embryo－Dissection－and－Micromanipulation－Tools
Embryo Dissection and Micromanipulation ToolsHazel L. Sive, Robert M. Grainger, and Richard M. HarlandAdapted from "Equipment for Embryo Experiments,"
The－effects－ofdifferent－concentrations－oflithium－chloride－on－thedevelopment
AbstractPrevious studies by Stachel and colleagues indicate that lithium chloride induction of post-midblastular Brachydanio rerio embryos results in
SDS－Whole－Cell－Extracts
-Use sterile technique and sterile solutions in steps 1 to 3.-1. Using a saturated starter culture, inoculate 25 to 30 ml of appropriate media in a 12
细胞凋亡（TUNEL，TUNEL染色）检测技术
细胞凋亡是指为维持内环境稳定， 生物体内细胞在特定的内源或外源信号诱导下，其死亡途径被激活，并在有关基因的调控下发生的程序性死亡过程。细胞凋亡是程序性死亡过程的一种主要形式，它涉及染色质凝聚和外周化、细胞质减少、核片段化、细胞质致密化、与周围细胞联系中断、内质网与细胞膜融合，最终细胞片段化形成许多
Isolation－of－mouse－embryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome
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
Isolation－of－Zebrafish－embryos
Zebrafish will mate and deposit fertilized eggs on the bottom of the tank at 'dawn'. They can be accustomed to lay at any convenient time by k