fluorescein Protocols

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

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

Protocol for fluorescein labeling of proteins. - [Read Fluorescein Labeling of Proteins Protocol]

Category: Molecular Imaging: Immunofluorescence Microscopy

Fluorochrome Absorption and Emission Spectra. BD Biosciences. Includes spectra of Allophycocyanin (APC), BD Cy-Chrome, Fluorescein isothiocyanate (FITC), R-phycoerythrin (R-PE), PE-Texas Redâ„¢, Peridinin chlorophyll protein (PerCP), PerCP-Cy5.5, APC-Cy7, Texas Redâ„¢. - [Read Fluorochrome Absorption and Emission Spectra]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

This simple cell culture-based cytotoxicity test (in which cell viability is determined by uptake of the dyes ethidium bromide and fluorescein acetate) has been developed as a general test for acute toxicity. - [Read LS-L929 Cytotoxcitiy Test]

Category: PCR Protocols: Real-Time PCR Protocols

In multiplex real-time PCR, different sets of primers with different labels are used to amplify separate genes from the template DNA in one tube.  This protocol uses LUX (Light Upon eXtension) primers from invitrogen.  FAM (6-carboxy-fluorescein) is used to label the gene of interest, and JOE (6-carboxy-4', 5'-dichloro-2',7'-dimethoxy-fluorescein) is used to label a housekeeping gene as an internal control to normalize between different reactions. - [Read Multiplex Real-Time PCR Protocol]

Category: Nucleic Acid Modification Protocols: Nucleic Acid Labeling Protocols

Protocol for nick-translation labeling of ds DNA with nick translation mixes for in situ probes. Includes: Labeling reaction with DIG-dUTP or Biotin-dUTP; Labeling reaction with Fluorescein-dUTP, or Tetramethylrhodamine-dUTP; Purification of labeled probe. - [Read Nick-Translation Labeling of ds DNA with Nick Translation Mixes for In Situ Probes Protocol]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol for oligonucleotide tailing with a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP mixture.
- [Read Oligonucleotide Tailing with a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP Mixture Protocol]

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

Protocol for oligonucleotide tailing with a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP mixture. - [Read Oligonucleotide Tailing With a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP Mixture Protocol]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol for PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. Includes: PCR DIG labeling reaction for moderately labeled probes; PCR fluorescein labeling reaction for direct in situ probes.
- [Read PCR Labeling of Ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes]

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

Protocol for PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. Includes: PCR DIG labeling reaction for highly labeled probes containing unique sequences; PCR DIG labeling reaction for moderately labeled probes; PCR fluorescein labeling reaction for direct in situ probes. - [Read PCR Labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes Protocol]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

This protocol describes a method for observing and measuring the movement of RNA molecules in the nucleus of living mammalian cells. Caged fluorescein-labeled DNA oligonucleotides are introduced into living mammalian cells, where they demonstrably hybridize to complementary RNA. After site-specific photoactivation at desired sites within the cell, the RNA movements away from those sites are followed and digitally recorded using a rapid acquisition microscopy system. - [Read Photoactivation-Based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, biotin, or fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotin- or Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

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

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, Biotin, or Fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotinor Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

Category: PCR Protocols: Real-Time PCR Protocols

Protocol uses FAM-(6-carboxy-fluorescein) or JOE-(6-carboxy-4', 5' -dichloro-2',7' -dimethoxy-fluorescein) labeled LUX (Light Upon eXtension) primers, which can quantify 100 or fewer copies of the target DNA in a background of nonspecific templates, over a broad dynamic range of less than 100-107 copies.  It uses uracil deglycosylase (UDG) to minimize the risk of carryover contamination, and includes a melting curve analysis of the product. - [Read Real-Time PCR Protocol]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol for RNA labeling by in vitro transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix. - [Read RNA Labeling by In Vitro Transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix]

Category: Cytogenetics Protocols: RNA In Situ Hybridization Protocols

Protocol for RNA labeling by in vitro transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix. A PCR fragment that has the appropriate promoter ligated to its 5’-ends can also serve as a transcription template. The procedure described incorporates one modified nucleotide (DIG-, Biotin-, or Fluorescein-UTP) at approximately every 20 – 25th position in the transcripts. - [Read RNA Labeling by In Vitro Transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix]

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

Protocol for RNA labeling by in vitro transcription of DNA with DIG, biotin or fluorescein RNA labeling mix. - [Read RNA Labeling by In Vitro Transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix]