techniques Protocols

Category: Antibody Protocols: Antibody Labeling Protocols

This protocol describes a simple chemical oxidation method for labeling antibodies with iodine. Iodide-125 (supplied as NaI) is oxidized to form iodine-125 (I2), which attacks tyrosyl and histidyl side chains. The iodinated antibodies are easily detected and quantitated using gamma counters or film. They are used primarily in immunoassays, but other techniques can be adapted conveniently to the iodine detection method. - [Read Labeling Antibodies with Iodine Protocol]

Category: Antibody Protocols: Antibody Labeling Protocols

This method for tagging monoclonal antibodies involves growing hybridomas in the presence of radioactive amino acids. This protocol can be particularly useful when conventional labeling techniques cause the antibody to lose activity. The labeled antibodies that result are essentially identical to the unlabeled antibodies. - [Read Labeling Monoclonal Antibodies by Biosynthesis Protocol]

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

LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, etc. Protocol provides a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1–1.5 h. - [Read Laser-capture Microdissection Protocol]

Category: Molecular Imaging: Light Microscopy

Ahmanson Center for Advanced Electron Microscopy and Imaging. Use of Semi-Thin Cryosections for Light Microscopy, Immunolabeling of Cryosections on Glass Slides, Problems with Autofluorescence. House Ear Institute - [Read Light Microscopy Techniques]

Category: Microscopy Protocols: In Vivo Imaging Protocols

Caenorhabditis elegans, a small (adults are ~1 mm long), free-living soil nematode that feeds on bacteria, is an ideal organism for applying various live microscopy techniques. This protocol describes useful techniques for preparing C. elegans for live microscopic analysis. Details of sample preparation depend on the developmental stage of the worm to be studied. - [Read Live Imaging of Caenorhabditis elegans: Preparation of Samples]

Category: C. elegans Protocols

Protocol describes useful techniques for preparing C. elegans for live microscopic analysis. Details of sample preparation depend on the developmental stage of the worm to be studied. - [Read Live Imaging of Caenorhabditis elegans: Preparation of Samples]

Category: RNA Protocols: RNA-binding Protein Purification

Magnetic techniques for the isolation and purification of proteins and peptides. Safarik et al. 2004 - [Read Magnetic techniques for the isolation and purification of proteins and peptides]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Live-cell imaging techniques provide a critical insight into the fundamental nature of cellular and tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein and synthetic fluorophore technology. Because of these advances, live-cell imaging has become a requisite analytical tool in most cell biology laboratories. - [Read Maintaining Live Cells on the Microscope Stage]

Category: Primary Cell Isolation and Culture Protocols

There are several manual methods that can be used to perform tissue microdissection. Techniques using hand-held tools as well as mechanical micromanipulator-based approaches have been described. However, speed and precision are the most important parameters and any method that achieves these is adequate. Investigators should also expect to invest time initially by practicing on 10 to 20 cases to begin to feel comfortable with the technique. - [Read Manual Microdissection]

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

Human tissues are comprised of multiple interacting cell populations in a complex three dimensional arrangement with each cellular phenotype determined by a unique profile of mRNA and protein expression. Before microdissection techniques were developed, the only analysis tools for phenotypic studies were primarily immunohistochemistry and in-situ hybridization. While useful, these tools are limited to single gene analysis and, in general, do not allow qualitative studies. - [Read Microdissection Overview]

Category: Plant Biology Protocols

The standard protocol for in situ hybridizations in plants still involves fixing fresh tissue, embedding the tissue in wax, sectioning with a microtome and detection of the transcripts of interest using labeled RNA-probes. This protocol concentrates only on nonradioactive methods, as they are easy to perform, very sensitive and even faster than techniques involving radioisotope labels. - [Read Molecular and Biochemical Analysis of Arabidopsis Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Mouse ES Cell Culture Techniques. List of protocols. The Bradley Lab. - [Read Mouse ES Cell Culture Techniques]

Category: Microscopy Protocols: Optical Microscopy Protocols

Diffraction-limited optical microscopy requires that the spatial resolution of an image is limited by the wavelength of the incident light & by the numerical apertures of the condenser & objective lens systems.The development of near-field scanning optical microscopy (scanning near-field optical microscopy) has allowed for a imaging technique that retains the various contrast mechanisms afforded by optical microscopy methods while attaining spatial resolution beyond the optical diffraction limit - [Read Near-Field Scanning Optical Microscopy]

Category: Microscopy Protocols: Optical Microscopy Protocols

Near-field scanning optical microscopy can achieve spatial resolution performance beyond the classical diffraction limit by employing a sub-wavelength light source or detector positioned in close proximity to a specimen. Such a sub-wavelength source usually consists of an aperture at the end of a tapered probe, which functions basically as a wave guide. Includes info.: Fiber Probe Fabrication; Pulling Method; Meniscus Etching; Selective Etching; Apertureless and Alternative Probe Designs etc. - [Read Near-Field Scanning Optical Microscopy: NSOM Probes]

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Materials for cresyl violet staining protocol (see methods section) ... Histology also refers to the techniques used to prepare tissue for microscopic study ... Neuroscience Histology Methods - [Read Neuroscience Histology Methods]

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

Quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Other PCR Procedures. - [Read Older RT-PCR and PCR Techniques Page.]

Category: PCR Protocols

Quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Other PCR Procedures. - [Read Older RT-PCR and PCR Techniques Page.]

Category: Molecular Imaging

Optimal Microscopy Primer - Molecular Expressions. Introduces most microscopy techniques - [Read Optimal Microscopy Primer - Molecular Expressions]

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: PCR Optimization Protocols

PCR PRIMER DESIGN AND REACTION OPTIMISATION: Molecular Biology Techniques Manual - Rybicki - [Read PCR PRIMER DESIGN AND REACTION OPTIMISATION: Molecular Biology Techniques Manual]

Category: PCR Protocols: PCR Optimization

PCR PRIMER DESIGN AND REACTION OPTIMISATION: Molecular Biology Techniques Manual - Rybicki - [Read PCR PRIMER DESIGN AND REACTION OPTIMISATION: Molecular Biology Techniques Manual]

Category: Plant Biology Protocols: Plant Tissue Culture

Plant tissue culture techniques. Pages 151-174, in Tested studies for laboratory ... engineering protocols employ callus initiation procedures after DNA has ... Mineo Zoology Univ. Toronto - [Read Plant Tissue Culture Techniques]

Category: Plant Biology Protocols: Arabidopsis Protein Peptide Protocols

Coimmunoprecipitation is one of the most useful techniques for revealing protein-protein interactions. Good negative controls to verify the specificity of the coimmunoprecipitation procedure are (1) performing the same immunoprecipitation experiment using beads coupled to the preimmune serum and (2) probing the Western blot with antibodies against protein known not to interact with the coimmunoprecipitated proteins under physiological conditions. - [Read Protein Coimmunoprecipitation in Arabidopsis Protocol]

Category: Protein Protocols: Protein Expression Protocols

Protein Expression Techniques Advantages and Disadvantages. Molecular Station. - [Read Protein Expression Techniques Advantages and Disadvantages]

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

Cryopreserved PBMCs are a common specimen source for studies of immunological responses to vaccines, immunotherapies, etc. The health and viability of cells recovered post-cryopreservation are of course critical to the success and accuracy of immunological assays performed on them. This protocol standardizes PBMC isolation and cryopreservation techniques, specifically for the assessment of thawed cells by cytokine flow cytometry. - [Read Protocol for Isolation, Cryopreservation, and Thawing of PBMCs]

Category: RNA Protocols: RT-PCR and cDNA synthesis

REVERSE TRANSCRIPTION PCR: Molecular Biology Techniques Manual - Rybicki - [Read REVERSE TRANSCRIPTION PCR: Molecular Biology Techniques Manual]