primary Protocols

Category: Cloning Protocols: Baculovirus Protein Expression Protocols

Protocol for amplification of Baculovirus plaques and preparation of high titer virus stocks. Includes: Primary Amplification; Secondary Amplification. - [Read Amplification of Baculovirus Plaques and Preparation of High Titer Virus Stocks Protocol]

Category: Antibody Protocols: Antibody Labeling Protocols

Once tissues are fixed and permeabilized, the antibodies are added. These antibodies can be labeled directly or detected by a labeled secondary reagent. For indirect detection, any reagent that binds specifically to the primary antibody can be "tagged" and used to locate the antibody. The possible reagents include anti-immunoglobulin antibodies, protein A or G, or, if the first antibody is labeled with biotin, streptavidin. They can be labeled with enzymes or gold. - [Read Binding Antibodies to Tissue Sections Protocol]

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocols for blocking solutions. Includes: Endogenous peroxidase blocking solution (H2O2-Azide); Non-specific antibody background blocking solution: swine serum; Non-specific antibody background blocking solution: MILK; Enzymatic antigen retrieval solution; Unconjugated, biotin- and fluorochrome-conjugated antibody (primary) solutions; Unconjugated, biotin- and fluorochrome-conjugated antibody (secondary) solutions; Enzyme-conjugated secondary antibody solutions.; etc.. - [Read Blocking Solutions Protocols]

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Detailed Neuronal cell transfection protocol and methods. Steven Finkbeiner. UCSF. - [Read Calcium Phosphate Transfection of Neurons in Primary Culture]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for CD90 (Thy-1) antibody staining for Immunohistochemistry. Includes: Primary Antibody; Antigen Retrieval; Detection Methods; Chromogen Substrate; Counterstain. - [Read CD90 (Thy-1) Antibody Staining Protocol for Immunohistochemistry]

Category: Molecular Imaging: Immunofluorescence Microscopy

Cell Staining for Immunofluorescence Microscopy. Includes protocols for fixing the cells, Coverslip Preparation for Adherent Cells, Coverslip Preparation for Non-Adherent Cells, Paraformaldehyde Fixation, and Methanol/Acetone Fixation. Blocking protocols include blocking with primary antibody, and incubation with secondary antibody. - [Read Cell Staining for Immunofluorescence Microscopy]

Category: Microscopy Protocols: Confocal Microscopy Protocols

Includes staining protocols: Triple-Staining Adherent Cells with MitoTracker, Phalloidin,& Nuclear Dyes; Staining Adherent Cells with Cytokeratin Primary Antibodies & Synthetic Fluorophores; Staining Adherent Cells with Intermediate Filament Primary Antibodies & Synthetic Fluorophores; Staining Cells & Tissue Cryosections with Tubulin Primary Antibodies, Phallotoxins,& Synthetic Fluorophores; Triple-Staining Tissue Cryosections with Wheat Germ Agglutinin, Phalloidin,& Nuclear Dyes; etc. - [Read Confocal Microscopy Specimen Preparation Using Synthetic Fluorophores and Immunofluorescence]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

Derivation of Mouse Embryonic Fibroblast Primary Cultures. MEF. MEFP - [Read Derivation of Mouse Embryonic Fibroblast Primary Cultures DOC]

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: 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: ELISA Protocols

Direct Elisa Using Primary Antibody Protocol. Abcam. - [Read Direct Elisa Using Primary Antibody Protocol]

Category: Immunology: T Cell Protocols

Protocol for Dlgh1 knockdown in primary OT-1 T lymphocytes. - [Read Dlgh1 Knockdown in Primary OT-1 T Lymphocytes Protocol]

Category: Transfection Protocols

Transfection protocol used in the Dlgh1 knockdown in primary OT-1 T lymphocytes. - [Read Dlgh1 Knockdown in Primary OT-1 T Lymphocytes: Transfection Protocol]

Category: Molecular Imaging: Electron Microscopy Protocols

E.M. processing schedule - epoxy resin E.M. processing schedule - acrylic resin Primary fixative and buffers Veterinary Pathology. ELECTRON MICROSCOPY (EM) protocols and methods - [Read ELECTRON MICROSCOPY (EM) protocols and methods]

Category: Molecular Imaging: Electron Microscopy Protocols

The protocol for immunolabeling for electron microscopy depends mostly on the primary antibody and its ability to recognize antigen under particular ...Electron Microscopy of the Cytoskeleton of Cultured Cells. Boris Lab. - [Read Electron Microscopy of the Cytoskeleton of Cultured Cells]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Growth of Primary Cell Culture and Viral Vector Handling Protocols]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

Category: Cell Biology and Cell Culture: Hepatocyte Cell Culture

Primary culture of hepatocytes is an in vitro model widely used to ... protocols for cytochromes P450IIIA in vivo and in primary cultures of animal and ...Human Hepatocyte Culture Protocol - [Read Human Hepatocyte Culture Protocol]

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol describes the direct detection of RNA on DNA microarrays using Hybrid Capture (HC) technology and the HC ExpressArray Kit developed by Diagene.  The kit uses a proprietary antibody that binds specifically to RNA:DNA hybrids and a second, fluorescently labeled, antibody that detects the primary antibody.  Total RNA is applied directly to a glass-spotted DNA microarray, and stable RNA:DNA hybrids are visualized via a Cy3-labeled secondary antibody. - [Read Hybridization and Detection Using the HC ExpressArray Kit 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: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for Immunoblot. Includes: Staining and Laser Capture Microdissection; Protein Separation by Polyacrylamide Gel Electrophoresis; Electrophoretic Transfer To a Membrane (Nylon, PVDF or Nitrocellulose); Primary and Secondary Antibody Incubations; Visualization. - [Read Immunoblot Protocol]

Category: Immunology: Immunofluorescence Protocols

Protocol for immunofluorescence labeling of cells. Includes: Cell Preparation; Fixation; Application of Primary Antibody; Application of Secondary Antibody and Evaulation. - [Read Immunofluorescence Labeling of Cells 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]