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DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_124-126.pdf

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol describes here a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. Includes: In situ hybridization with DIG-labeled probes; Detection of DIG-labeled probes; Fluorescence microscopy. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System]

DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_124-126.pdf

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol describes a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. This procedure can be used to detect single copy sequences as small as 1 kb on human metaphase chromosomes. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol]

DNA Sequencing Methods - http://www.genome.ou.edu/protocol_book/protocol_partIV.html#IV.C

Category: DNA Protocols: DNA Sequencing Protocols

Methods for DNA sequencing. Includes: Bst-catalyzed radiolabeled DNA sequencing; Radiolabeled sequencing gel preparation, loading, and electrophoresis; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye primers; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye terminator reactions; Sequenase[TM] catalyzed sequencing with dye-labeled terminators; etc. - [Read DNA Sequencing Methods]

Drosophila Non-fluorescent Protein and RNA Double-Labeling Protocol - http://www.utoronto.ca/krause/double%20labeling.html

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol for Drosophila non-fluorescent protein and RNA double-labeling. - [Read Drosophila Non-fluorescent Protein and RNA Double-Labeling Protocol]

Extraction and Purification of RNA from Blood Samples for Fluorescent mRNA Differential Display - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4125

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols: RNA Isolation fromTissues or Cultured Cells Protocols

Protocol is the first in a set of three describing fluorescent mRNA differential display (FDD or FDDRT-PCR). The method begins with the harvesting of total RNA from blood samples. - [Read Extraction and Purification of RNA from Blood Samples for Fluorescent mRNA Differential Display]

Extraction and Purification of RNA from Tissue Samples for Fluorescent mRNA Differential Display - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4124

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols: RNA Isolation fromTissues or Cultured Cells Protocols

Protocol is the first in a set of three describing fluorescent mRNA differential display (FDD or FDDRT-PCR). The method begins with the harvesting of total RNA from tissue samples. - [Read Extraction and Purification of RNA from Tissue Samples for Fluorescent mRNA Differential Display]

Extraction and Purification of RNA from Tissue-Cultured Cells for Fluorescent mRNA Differential - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4123

Category: PCR Protocols

Protocol is the first in a set of three describing fluorescent mRNA differential display (FDD or FDDRT-PCR). The method begins with the harvesting of total RNA from the tissue-cultured cells of interest. For other starting materials, such as blood samples, please see Extraction and Purification of RNA from Blood Samples for Fluorescent mRNA Differential Display. - [Read Extraction and Purification of RNA from Tissue-Cultured Cells for Fluorescent mRNA Differential]

Flow Cytometry Analysis Protocol - http://www.cbrinstitute.org/labs/springer/protocols/qing_facs.html

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

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining and Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Flow Cytometry Analysis Protocol - http://www.cbrinstitute.org/labs/springer/protocols/qing_facs.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining; Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Fluorescence In Situ Hybridization (FISH) for DNA Replication Origins Protocol - http://www.natureprotocols.com/2006/10/27/fluorescence_in_situ_hybridiza.php

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for fluorescence in situ hybridization (FISH) for DNA replication origins. Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique used for the detection of specific chromosomal rearrangements and applicable to many different specimen types. FISH is widely used for several diagnostic applications. - [Read Fluorescence In Situ Hybridization (FISH) for DNA Replication Origins Protocol]

Fluorescent Dye Properties Data - http://www.sigmaaldrich.com/img/assets/8181/FluorescDyeProp.pdf

Category: Immunology: Immunofluorescence Protocols

Data for fluorescent dye properties. Includes: Fluorochrome, Excitation in(nm), Emission in (nm) and Color Application. - [Read Fluorescent Dye Properties Data]

Fluorescent Dyes for Flow Cytometric Analysis - http://www.ebioscience.com/ebioscience/appls/Dyes.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Fluorescent dyes absorb light at certain wavelengths and in turn emit their fluorescence energy at a higher wavelength. Each dye has a distinct emission spectrum, which can be exploited for multicolor analysis. eBioscience antibodies are available conjugated to a wide variety of fluorochromes. - [Read Fluorescent Dyes for Flow Cytometric Analysis]

Fluorescent In Situ Transcription in Cells and Tissues Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6639A7FEDD7A49BBC3758516F99557&objectid=6677C9E8B913BFB72E9F654025F3C39A

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

Fluorescent indicators for imaging protein phosphorylation in single living cells - http://www.nature.com/nbt/journal/v20/n3/full/nbt0302-287.html

Category: Protein Protocols: Protein Phosphorylation Protocols

Fluorescent indicators for imaging protein phosphorylation in single living cells. Using Phocuses, genetically encoded fluorescent indicators, one can visualize signal transduction based on protein phosphorylation in living cells. Moritoshi Sato1, Takeaki Ozawa1, Kouichi Inukai2, Tomoichiro Asano2 & Yoshio Umezawa1. Nature Biotechnology - [Read Fluorescent indicators for imaging protein phosphorylation in single living cells]

Fluorescent Reagents for Live Cell Imaging and Their Introduction into Cells - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.ip20

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Most biological specimens are relatively transparent, so details of internal and intracellular morphology are difficult to image in untreated living specimens using simple bright-field techniques. Fluorescence microscopy offers greater advantages and possibilities for increasing contrast and determining the specific localization of molecules in cells. Article outlines the three methods most commonly used to introduce an appropriate label into Drosophila tissue without perturbing the process. - [Read Fluorescent Reagents for Live Cell Imaging and Their Introduction into Cells]

Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4439

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Protocol provides a method for analysis of cell surface antigens using direct immunofluorescence, i.e., when the antibody being used is directly labeled to a fluorescent dye. After staining, the cells can be analyzed immediately by flow cytometry or fixed and stored for later analysis. - [Read Fluorescent Staining of Cell Surface Antigens on Viable Cells Using Direct Immunofluorescence]

Fluorescent Staining of Cells - http://ylwang.umassmed.edu/protocol/cfs/stain.htm

Category: Molecular Imaging: Immunofluorescence Microscopy

Fluorescent Staining of Cells protocol. Yu-li Wang Lab. Fluorescent Phalloidin Staining, Immunofluorescence Staining and procedures for staining. - [Read Fluorescent Staining of Cells]

Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4458

Category: Nucleic Acid Detection Protocols

Protocol describes the quantitation of DNA using Hoechst 33258, a fluorescent dye that binds to double-stranded DNA. Fluorometry is simple and more sensitive than spectrophotometry, and allows the detection of nanogram quantities of DNA. The assay can only be used to measure the concentration of DNAs whose sizes exceed ~1 kb, as Hoechst 33258 binds poorly to smaller DNA fragments. - [Read Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol]

Human DNA Microarray Hybridization - http://cmgm.stanford.edu/pbrown/protocols/5_hyb_human.html

Category: DNA Microarray Protocols

Preparation of Fluorescent DNA Probe from HUMAN mRNA or Total RNA using Direct Incorporation Washing and Scanning Arrays. Brown Lab. - [Read Human DNA Microarray Hybridization]

Hybridization Protocol for Hybridizing fluorescent cDNA to slide - http://research.nhgri.nih.gov/microarray/hybridization.shtml

Category: DNA Microarray Protocols

Hybridization Protocol for Hybridizing fluorescent cDNA to slide. - [Read Hybridization Protocol for Hybridizing fluorescent cDNA to slide]

Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4673

Category: Reporter Gene Assays: GFP Assay Protocols

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. These methods should be followed in the order presented, but any of them can be omitted when not needed. - [Read Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle]

Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4673

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]

Immunofluorescent Staining for Flow Cytometry Protocols - http://www.ebioscience.com/ebioscience/appls/FCS.htm

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]

Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis - http://www.bdbiosciences.com/pharmingen/protocols/Intracellular_Cytokines.shtml

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Fluorescent anti-cytokine and monoclonal antibodies are useful for the intracellular staining and multiparameter flow cytometric analysis of individual cytokine-producing cells within mixed cell populations. BD Biosciences. - [Read Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis]

Improvements in Fluorescent In Situ RT-PCR for Medical and Research Use - http://iurtc.iu.edu/ott/technol/techs/0007.htm

Category: PCR Protocols: In-situ PCR Protocols

Improvements in Fluorescent In Situ RT-PCR for Medical and Research Use. Robert Bacallao, M.D. et al - [Read Improvements in Fluorescent In Situ RT-PCR for Medical and Research Use]

Articles (1-3 of 3)

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.