powerful Protocols

Category: Antibody Microarray Protocols

Antibody Microarrays Are a Powerful Tool for Proteomics. Genomics and Proteomics - [Read Antibody Microarrays Are a Powerful Tool for Proteomics]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Sorting Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Describes the standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors]

Category: Stem Cell Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Here, we described our standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells and Myeloid Progenitors and Isolating RNA]

Category: Immunology: Immunoprecipitation Protocols: Immunoprecipitation of Radiolabeled Cells Protocols

Most powerful and convincing method to determine if a specific protein is phosphorylated in a physiologically relevant manner is to assay phosphorylation in situ. The procedure described involves incubating cultured cells (e.g., primary neuronal cultures or transfected cells) or tissue preparations (e.g., hippocampal slices) with [32P]orthophosphate, which is then taken up by the cells or tissues and incorporated into the γ-phosphate position of ATP. - [Read Detection of Protein Phosphorylation in Tissues and Cells Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

DNA is best isolated from bacteriophage lambda particles by digesting the viral coat proteins with a powerful protease such as proteinase K, followed by extraction with phenol:chloroform. This procedure is easily adapted for use with smaller-scale preparations of bacteriophage lambda. - [Read Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Proteinase K and SDS Protocol]

Category: Epigenetics and Methylation Protocols: Bisulfite Treatment Methylation Assay Protocol

Hairpin-bisulfite PCR will be a powerful tool in studying other processes for which genetic or epigenetic information differs on the two complementary strands of DNA. Laird et al., 2003. PNAS - [Read Hairpin-bisulfite PCR: Assessing epigenetic]

Category: Microinjection Protocols

Fluorescence microscopy provides a powerful tool for imaging molecular components in living cells. Specific molecular components can be efficiently labeled by a combination of three methods: chemical transfection of GFP-fusion constructs, staining of chromosomes with the DNA-specific, fluorescent dye Hoechst 33342, and microinjection of fluorescently conjugated proteins. This procedure provides an example of using all three methods in sequence to label components of living HeLa cells. - [Read Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle]

Category: Microscopy Protocols

The atomic force microscope (AFM) is one of the most powerful tools for determining the surface topography of native biomolecules at subnanometer resolution. The AFM can also provide insight into the binding properties of biological systems. In order to determine the specific interaction between two kinds of molecules (e.g., avidin and biotin). Includes information on principle of AFM and application of AFM. - [Read Imaging, Measuring and Manipulating Native Biomolecular Systems with the Atomic Force Microscope]

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: Microscopy Protocols: Fluorescence Microscopy Protocols

Multiphoton fluorescence microscopy is a powerful new technology that enables the acquisition of optical sections without the use of a pinhole aperture typically used for confocal microscopy. The technique is based upon the two-photon principle: A fluorescent molecule simultaneously absorbs two photons producing an electronic transition from the ground to excited state equal to two times the energy of each incident photon. - [Read Multiphoton Images from LSM 510 NLO System]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

A powerful way to identify a mutation in the gene of interest and to test mutant plants for phenotypes that are predicted to result from loss of function of that gene is by PCR screening. Pools of insertion lines are screened using one primer corresponding to the gene of interest and one primer corresponding to the end of the insertion element. The synthesis of a product indicates the presence of an insertion in the gene of interest. - [Read Screening DNA Pools for T-DNA Insertions in Arabidopsis Genes Protocol]

Category: Genetics and Genomics: Cancer Genetics Protocols: Single Strand Conformational Polymorphism(SSCP) Protocols

Single-strand confirmation polymorphism analysis (SSCP) is a powerful and robust method for the detection of DNA sequence changes (single-base substitutions) based on shifts in electrophoretic mobility. In this protocol, the target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. - [Read Single-Strand Conformation Polymorphism Analysis Protocol]

Category: Neuroscience Protocols: Patch Clamping Protocols

The cell-attached capacitance recording technique is a powerful technique that has been successfully used to resolve single vesicle fusion and fusion pore conductance.  This technique, however, has not been applied to synapses because of the difficulty in accessing release sites.  Here, we developed a technique to expose release sites in a large nerve terminal, the calyx of Held, which contains clear-core glutamatergic vesicles. - [Read The Cell Attached Capacitance Recording Technique]

Category: Electrophysiology Ion Channels

The Whole-Cell Patch-Clamp Technique – a powerful tool to approach CFTR function. Martin J. Hug. University of Muenster, Institute of Physiology. Germany - [Read The Whole-Cell Patch-Clamp Technique – a powerful tool to approach CFTR function]