tissue Protocols

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for histochemical staining. This protocol has been optimized for b-galactosidase and human placental alkaline phosphatase staining in retinal tissue and cultured cells. - [Read Histochemical Staining Protocol]

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Protocols for cell fixation and tissue fixation. HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block techniques. Embedding Tissue Into Frozen Blocks. Fixing Cells with Fresh Paraformaldehyde. Preparation of Frozen Sections. Immunohistochemistry. Paraffin sections and Frozen sections. DERC Univ. Colorado PDF. - [Read HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block PDF]

Category: Signalling Signal Transduction Protocols

This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography. Includes information: H1 Kinase Assay on Individual Xenopus Oocytes; H1 Kinase Assay on Xenopus Egg Extract Samples; H1 Kinase Assay on Tissue Culture Cells; Helpful protocol hints. - [Read Histone H1 Kinase Activity Assay]

Category: Histology Protocols

Histotechniques - Manual. Great Histology Protocol Handbook. Includes: Tissue Processing, Specimen Accessioning, Fixation, Fixatives, Tissue Processing, Sectioning, H and E staining, more. - [Read Histotechniques - Manual]

Category: Immunohistochemistry Protocols: Fixation Protocols

Information on histotechniques. Includes: Fixation - types of fixatives; factors affecting fixation; General usage of fixatives; Tissue Processing; Sectioning; Frozen Sections; Staining; H and E staining; Coverslipping; Decalcification; Artefacts in Histologic Sections; Problems in Tissue Processing. - [Read Histotechniques Fixation]

Category: Cytoskeleton Protocols: Actin Myosin Protocols

The same GFP-tagged actin construct used in cell transfection experiments has been used to produce transgenic mice. Transgenic animals allow the imaging of brain tissue in the intact animal, as acutely cut slices or as organotypic slice cultures. They also serve as a source of cells for imaging neurons at high resolution in dispersed low-density cell culture. In contrast to cells transfected in culture, where the level of actin-GFP expression in neurons varies considerably, transgenic mice... - [Read Imaging Actin in Tissue Slices from Transgenic Mouse Brain Protocol]

Category: Microscopy Protocols: Electron Microscopy Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope]

Category: Plant Biology Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope Protocol]

Category: Cell Culture Protocols: Cell Immortalization Protocols

Cultivating animal cells in the laboratory is an indispensable technique for cell biologists. However, most normal primary cell lines, while faithfully reproducing the phenotype of their tissue of origin, do not grow indefinitely in culture. After a series of population doublings (the number of which varies by species, cell type, and culture conditions) primary cells enter a state where they no longer divide. - [Read Immortalization of Cells in Culture]

Category: Western Blot Protocols: General Western Blot Methods Protocols

For immunoblotting experiments, it is often important to compare the total amount of an antigen from many different sources or to learn if a particular source has the antigen under study.  In the approach described here, tissue cultures, bacteria, yeast cells, tissues, and other sources of antigens are disrupted directly in an electrophoresis sample . - [Read Immunoblotting: Preparing Cell Lysates Protocol]

Category: Immunology: Immunocytochemistry Protocols

Using anti-BrdU, Diaminobenzadine (DAB) brown detection. Includes counterstaining protocol. - [Read Immunocytochemical Identification of Proliferating Cells In Vascular Tissue]

Category: Molecular Imaging: Immunofluorescence Microscopy

B or T cells in suspension, adherent cells on chambered coverglass or chamberslides, cryostat sections of unfixed, OCT embedded tissue. Susan Anderson. - [Read Immunofluorescence / confocal microscopy]

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of the cell. Antibody molecules for a specific target molecule are exposed to the cell or tissue being investigated. The binding of these molecules is detected by incubating the sample with a secondary antibody specific for immunoglobulin molecules and conjugated to fluorophore. - [Read Immunofluorescence Staining Protocol]

Category: Immunology: Immunofluorescence Protocols

Protocol for immunofluorescence staining. Includes: Cell Lines; Frozen Sections; Paraffin Sections; Pretreatments of Tissue Sections. - [Read Immunofluorescence Staining Protocol]

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Protocols for immunofluorescent staining for flow cytometry. Includes: Protocol for Staining of Cell Suspensions of Murine Lymphoid Tissue; Protocol for Staining of Human Peripheral Blood; Fluorescent Dyes for Flow Cytometric Analysis - [Read Immunofluorescent Staining for Flow Cytometry Protocols]

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

Useful techniques to circumvent disruption of tissue structure in the analysis of gene expression are LCM and LDM. While they require specialized microscopes and systems, they are similar in that freshly-cut frozen tissue sections can be microdissected using either a general histological stain (like H&E) or by staining with fluorescently conjugated antibodies. The LCM system by Arcturus involves... - [Read Immunofluorescent Staining for the Laser Microdissection of Individual Cells Protocol]

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Protocol for immunofluorescent staining of cell suspensions of mouse lymphoid tissue. - [Read Immunofluorescent Staining of Cell Suspensions of Mouse Lymphoid Tissue Protocol]

Category: Immunohistochemistry Protocols

Protocol for immunohistochemistry enzyme HRP staining. Includes: Cell Linee; Frozen Sections; Paraffin Sections; Pretreatments of Tissue Sections. - [Read Immunohistochemistry Enzyme HRP Staining Protocol]

Category: Immunohistochemistry Protocols

Protocol describes the application of peroxidase or alkaline phosphatase conjugates in the immunohistochemical labeling of formalin-fixed, paraffin-embedded tissue sections. Includes: Removal of Paraffin and Rehydration; Antigen Retrieval - Unmasking of Antigen; Enzyme retrieval; Microwave retrieval; Inactivation of Endogenous Peroxidase; etc.. - [Read Immunohistochemistry Protocol]

Category: Molecular Biology Protocols: Carbohydrate Protocols

Indirect method measuring immunofluorescence coupled to second antibody. Best for membrane antigens in addition to intra- and extracellular antigens, may be applied to frozen tissue sections, to cells in suspension, and to cells attached to glass slides or coverslips. Tadashi Tai~Head, Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan - [Read Immunohistochemistry using Anti-Ganglioside Antibodies]

Category: Immunohistochemistry Protocols

Describes two methods for using the immunoperoxidase reaction to localize antigens at the electron microscope level; one for adherent cultured cells and one for tissue sections. The reaction conditions are first optimized at the light microscope level and then adapted for EM level observation. These methods allow for reliable detection of antigens at the cell surface, within the cell, and especially in membrane bounded organelles. - [Read Immunoperoxidase Methods for Localization of Antigens in Cultured Cells and Tissues]

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol for the immunoprecipitation (IP) of Homer 1a, injection of virions and in situ hybridization in the spinal cord. Includes: Immunoprecipitation (IP) of Homer1a from spinal cord; Injection of virions in the parenchyma of the spinal dorsal horn in vivo; Generation of cRNA probes; Analysis of DIG-dUTP incorporation; Tissue hybridization. - [Read Immunoprecipitation of Homer 1a, Injection of Virions and In Situ Hybridization in the Spinal Cord]

Category: Molecular Biology Protocols: Carbohydrate Protocols

Immunostaining thin layer chromatograms TLC is a very sensitive detection technique of functionally active carbohydrate ligands of protein receptors. Carbohydrate structures are detected in glycolipids from complex mixtures of molecules extracted from the relevant target tissue. Proteins analyzed can be antibodies, chimeric Ig proteins, selectins, lectins, toxins, and other carbohydrate binding proteins. John L. Magnani~GlycoTech Corporation, Rockville, Maryland - [Read Immunostaining Thin Layer Chromatograms Of Glycolipids]

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

Describes the basic principles of in situ hybridization and advantages and disadvantages of different methodologies that can be used. Includes: Probe Selection; Probe Generation; Probe Labels; Fixation of Tissue; Hybridization and Washing; Control Procedures. - [Read In situ Hybridization]

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for in situ hydbridization to tissue sections. Includes: Paraffin Sections; Frozen Sections; Probe preparation; Hybridization and washes; Autoradiography. - [Read In Situ Hybridization to Tissue Sections Protocol]