development Protocols

Category: Immunology: Chemotaxis

Chemotaxis Practical. Thierry Soldati. Department of Biological Sciences Animal and Plant Physiology. The assay in brief. Safety and Good Laboratory Practice: Working in a Cell Biology Laboratory. I-Chemotaxis Assay. Observation of Dictyostelium development stages. - [Read Chemotaxis Practical]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Method is for preparing chromosomes from single flower buds of A. thaliana. It does not kill the plants allowing the determination of their chromosome number throughout development. Includes: Preparations of Arabidopsis; Preparation of Chromosome; Staining Chromosomes. - [Read Chromosome Spreads from Flower Buds of Arabidopsis thaliana Protocol]

Category: Plant Biology Protocols

Protocol can be used for clearing intact non-ovule materials of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse root, seedling even flower development without sectioning. This protocol could also be used for clearing GUS stained material, after chlorophyll is removed by 70% ethanol. - [Read Clearing Arabidopsis Non-Ovule Materials With the HCG Solution Protocol]

Category: Plant Biology Protocols

Protocol can be used for clearing intact seeds of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse embryo and endosperm development without sectioning. - [Read Clearing Arabidopsis Ovule Materials with the Hoyer's Solution Protocol]

Category: Reporter Gene Assays: GFP Assay Protocols

The following protocol can be used for the development of stable cell lines expressing GFP fusion proteins. Although optimal transfection procedures (e.g., calcium phosphate, electroporation, or FuGENE 6 [Roche Applied Science]) vary depending on cell type, this general transfection procedure has been successful for stable transfection of HeLa, A-431, U2OS, BHK, and HT1080 cells. - [Read Constructing and Expressing GFP Fusion Proteins]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Culture and Characterization of Human Embryonic Stem Cells. DRAPER et al., STEM CELLS AND DEVELOPMENT 13:325Ð336 (2004) - [Read Culture and Characterization of Human Embryonic Stem Cells. PDF]

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for the development of techniques for primary culture of C. elegans embryonic neurons. - [Read Development of Techniques for Primary Culture of C. elegans Embryonic Neurons]

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons. Includes: Optimized cell culture protocol. - [Read Development of Techniques for Primary Culture of C. elegans Embryonic Neurons 2nd Part]

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons. - [Read Development of Techniques for Primary Culture of C. elegans Embryonic Neurons 3rd Part]

Category: Immunology: Immunoassays

Protocol for ELISA assay for NGF. Includes: ABSORPTION OF THE POLYCLONAL AND PREIMMUNE SERUM; BLOCKING; SAMPLE PREPARATION; PREPARATION OF NGF STANDARDS; PROTEIN RECOVERY; DESIGNING THE PLATE; APPLYING THE STANDARDS AND SAMPLES; APPLYING THE MONOCLONAL; APPLYING SECONDARY ANTIBODY; APPLYING STREPTAVIDIN; CHROMAGEN DEVELOPMENT; READING THE PLATE. - [Read Enzyme-Linked ImmunoSorbent Assay (ELISA) for NGF Protocol]

Category: Drosophila Protocols: Drosophila Genetics Protocols

FISH protocols for Drosophila. Includes: RNA Probe Preparation; Embryo Collection and Fixation; Single FISH on Drosophila embryos; Post-Fixation, Hybridization and Post-Hybridization Washes; Development of FISH Signal; Storage, Mounting and Viewing of Samples; Double FISH on Drosophila Embryos; RNA-Protein Double Labeling; FISH on Dissected Tissues. - [Read FISH Protocols for Drosophila]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Forward genetics is used to identify genes that are involved in particular biological processes. For example, genes required for disease resistance can be found by identifying mutants with reduced or increased disease resistance, genes that control flower development can be identified by searching for mutants with altered flower morphology, and genes encoding enzymes for tryptophan biosynthesis can be identified by searching for mutants that require exogenous tryptophan for growth. - [Read Forward Genetics in Arabidopsis: Finding Mutations that Cause Particular Phenotypes Protocol]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

After fixation, frozen sections are immunostained under RNase-free conditions using a rapid three-step streptavidin-biotin technique followed by dehydration. The immunostained sections are ready for LCM. Includes: Development of Immuno-LCM. - [Read Immuno-Laser Capture Microdissection Protocol]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

There are several strategies to visualize the antibody. For transmitted light microscopy, color development substrates for enzymes are often used. The antibody can be directly labeled with the enzyme. However, such a covalent link between an antibody and an enzyme might result in a loss of both enzyme and antibody activity. For these reasons several multistep staining procedures have been developed, where intermediate link antibodies are used. In this protocol use the Vectastain ABC-kit. - [Read Immunocytochemistry in Free-Floating Sections Protocol]

Category: Immunohistochemistry Protocols

Protocol for immunohistochemistry with AP-Conjugated (NBT/BCIP). Protocol extensively blocks slides, further diluting the primary antibody, lengthening the incubation and washing time, using a simple AP-conjugated secondary at high dilution and use a slow long development with the most powerful IHC development, NBT/BCIP. Includes: Single AP stainiing and Double AP staining. - [Read Immunohistochemistry with AP-Conjugated (NBT/BCIP) Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

Isolation of extraembryonic tissues allows one to study normal mouse development as well as the molecular basis of defects which cause fetal death. This protocol describes a method for isolating extraembryonic membranes from pregnant mice. - [Read Isolating Mice Extraembryonic Membranes Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolation of early neural-fold-stage embryos. - [Read Isolating Postimplantation Embryos: Early Neural-Fold-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause post-implantation defects. This protocol describes a method for isolating early primitive-streak-stage embryos. - [Read Isolating Postimplantation Embryos: Early Primitive-Streak-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating early somite-stage embryos (~8.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Early Somite-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating late primitive-streak-stage embryos (~7.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Late Primitive-Streak-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations that cause post-implantation defects. This protocol describes a method for isolating prestreak-stage embryos (~5.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Prestreak-Stage Protocol]

Category: Molecular Model Organisms: Mouse Animal Protocols: Mouse Genotyping Protocols

Contains the NHLBI Division of Intramural Research, including a list of labs, and the Office of Technology Transfer and Development. - [Read Laboratory of Developmental Biology, Division of Intramural Research, NHLBI, NIH]

Category: Lipid Protocols

Protocol for lipid analysis using thin layer chromotography. Protocol includes: Thin layer chromatography: Absorbent: Solvents, Development and Visualization. - [Read Lipid Analysis Using Thin Layer Chromotography Protocol]

Category: Neuroscience: Neuron Cell Culture Protocols

Neuronal Cell Culture PDF. Lesuisse and Martin. Wiley Periodicals 2002. - [Read Long-Term Culture of Mouse Cortical Neurons as a Model for Neuronal Development, Aging, and Death]

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

Measurement of Apoptosis and Other Forms of Cell Death. Jagan Muppidi, Melissa Porter, and Richard M. Siegel. As programmed cell death (PCD) or apoptosis has emerged as an important regulator of development and homeostasis in multicellular organisms, methods to quantify apoptosis and to distinguish it from necrosis have been developed. This unit presents a set of assays for these purposes, many of which are technically very simple and ideally suited to the study of hematopoietic cells. - [Read Measurement of Apoptosis and Other Forms of Cell Death]