biotin Protocols

Category: Cell Biology and Cell Culture: Apoptosis Protocols: TUNEL assay apoptosis

Apoptosis Detection Using Terminal Transferase and Biotin-16-dUTP (TUNEL -Enzyme Method). IHC World - [Read Apoptosis Detection Using TUNEL Enzymatic]

Category: Cell Biology and Cell Culture: Apoptosis Protocols: TUNEL assay apoptosis

Apoptosis Detection Using Terminal Transferase and Biotin-16-dUTP (TUNEL - Fluorescence Method). IHC World - [Read Apoptosis Detection Using TUNEL Fluorescence]

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol uses the BIOPRIME reaction kit from GibcoBRL to prepare biotin-labelled BAC DNA which is detected using FITC-Avidin (Vector Labs, DCS grade). Reagents from other manufacturers may work equally well but have not been tested. Includes: Labeling of BAC clones; Ethanol precipitation; Hybridization; Post-hybridisation treatment / detection. - [Read BAC-FISH Protocol]

Category: Mouse Protocols

Protocol describes the strongest way for lineage tracing on sections by using biotin dextran amine. - [Read BDA Mouse Lineage Tracing on Sections 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: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for BioPrime biotin labeling (FISH probe). - [Read BioPrime Biotin Labeling (FISH probe) Protocol]

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocol for blocking of unwanted non-specific staining. Includes: Blocking of endogenous enzymes; Blocking of endogenous fluorochromes; Blocking endogenous biotin; Blocking of endogenous Fc blocking; Blocking of crossreactive antigens in the tissue. - [Read Blocking of Unwanted Non-Specific Staining Protocol]

Category: Molecular Imaging: Immunofluorescence Microscopy

Blocking of unwanted non-specific staining in Immunofluorescence. Blocking of endogenous enzymes, Blocking of endogenous fluorochromes, Blocking endogenous biotin , Blocking of endogenous Fc blocking, Blocking of crossreactive antigens in the tissue. Cattoretti. Columbia University - [Read Blocking of unwanted non-specific staining in Immunofluorescence.]

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: Immunohistochemistry Protocols: Blocking Protocols

Protocol for blocking of unwanted non-specific staining. Includes: Blocking of endogenous enzymes; Blocking of endogenous fluorochromes; Blocking endogenous biotin; Blocking of endogenous Fc blocking; Blocking of cross reactive antigens in the tissue. - [Read Blocking Unwanted Non-Specific Staining Protocol]

Category: Cytogenetics Protocols

Protocol for CGH of biotin labeled DNA vs digoxigenin labeled DNA. Includes: Reprecipitate DNA's; Denature slides; Hybridization; Washing and staining. - [Read CGH of Biotin Labeled DNA vs Digoxigenin Labeled DNA Protocol]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

This protocol describes how to use DIG Chem-Link to directly label any DNA [e.g. plasmids, PCR products, cDNA prepared from mRNA] or RNA (e.g. total RNA, poly(A)+ mMRNA). The DIG Chem-Link or Biotin Chem-Link may also be used to label oligonucleotides. Includes: Required Purity of DIG Chem-Link Templates; Direct DIG Labeling of mRNA or cDNA with DIG Chem-Link; Key Product Required for Direct Labeling of DNA or RNA; Estimating the Yield of DIG-labeled Nucleic Acids. - [Read Chem-Link Labeling of DNA or RNA with DIG or Biotin Protocol]

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

Protocol for Chem-Link labeling of DNA or RNA with DIG or Biotin. Includes: Direct DIG Labeling of mRNA or cDNA with DIG Chem-Link; Estimating the Yield of DIG-labeled Nucleic Acids. - [Read Chem-Link Labeling of DNA or RNA with DIG or Biotin Protocol]

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

Detection of Phosphatidylserine Externalization During Apoptosis. Shailaja Kasibhatla et al. An early event in apoptosis is the externalization of phosphatidylserine (PS), a phospholipid normally restricted to the inner leaflet of the plasma membrane. This apoptotic event can be monitored using Annexin V, a PS-specific binding protein. This protocol uses Annexin V-FITC as a probe, but Annexin V-biotin is also available, and binding can be revealed using streptavidin-FITC or oth - [Read Detection Of Phosphatidylserine Externalization During Apoptosis (Subscription Required)]

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

The goal of this method is to identify transcriptionally active genes in cloned segments of genomic DNA. The protocol uses hybridization and affinity purification to recover biotin-labeled cDNAs that bind to a 500-kb segment of human DNA cloned in a BAC vector. However, the method can be easily adapted to other clones of genomic DNAs cloned in high-capacity vectors. - [Read Direct Selection of cDNAs with Large Genomic DNA Clones Protocol]

Category: Antibody Protocols: Epitope Mapping Antibody Protocols

A set of overlapping synthetic peptides is synthesized, each corresponding to a small segment of the linear sequence of a protein antigen and arrayed on a solid phase. The panel of solid-phase peptides is then probed with a test antibody, and bound antibody is detected using an enzyme-labeled secondary antibody. This method is very rapid and can be extraordinarily successful. - [Read Epitope Mapping Using Synthetic Biotin-Labeled Peptides Protocol]

Category: Microscopy Protocols

The atomic force microscope (AFM) is one of the most powerful tools for determining the surface topography of native biomolecules at subnanometer resolution. The AFM can also provide insight into the binding properties of biological systems. In order to determine the specific interaction between two kinds of molecules (e.g., avidin and biotin). Includes information on principle of AFM and application of AFM. - [Read Imaging, Measuring and Manipulating Native Biomolecular Systems with the Atomic Force Microscope]

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

After fixation, frozen sections are immunostained under RNase-free conditions using a rapid three-step streptavidin-biotin technique followed by dehydration. The immunostained sections are ready for LCM. Includes: Development of Immuno-LCM. - [Read Immuno-Laser Capture Microdissection Protocol]

Category: Immunohistochemistry Protocols

Protocol for immunohistochemistry on fixed, paraffin-embedded sections. This method is widely used and applies to the detection of the overwhelming majority of antigens, with few exceptions for which enzymatic retrieval is required. The method uses a strong chelating agent, EDTA. Includes: Double indirect AP; AP Developing solution; Indirect immunohistochemistry with avidin-biotin and HRP; HRP Developing solution. - [Read Immunohistochemistry on Fixed, Paraffin-Embedded Sections Protocol]

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with anti-phosphotyrosine:biotin conjugates. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation with Anti-Phosphotyrosine:Biotin Conjugates Protocol]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol for In situ hybridization to human metaphase chromosomes using DIG-, biotin-, or fluorochrome-labeled DNA probes and detection with fluorochrome conjugates. Includes: Pretreatment of metaphase spreads on slides; Denaturation and hybridization; Single color fluorescent detection with immunological amplification; Multicolor fluorescence in situ hybridization (Multicolor FISH); Results obtained with human metaphase chromosome spreads. - [Read In Situ Hybridization to Human Metaphase Chromosomes using DIG-, Biotin- or Fluorochrome-Labeled DNA]

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

Protocol for in situ hybridization to human metaphase chromosomes using DIG-, biotin-, or fluorochrome-labeled DNA probes and detection with fluorochrome conjugates. - [Read In Situ Hybridization to Human Metaphase Chromosomes Using DIG-, Biotin-, or Fluorochrome-Labeled]

Category: DNA Protocols: EMSA DNA-Protein Protocols

A non-radioactive electrophoretic mobility which works quite well using biotin and streptavidin detection. Non-specific and specific competititor, oligo labeling, Binding Reaction. Pierce - [Read Introduction to the EMSA (Gel Shift) Technique]

Category: Antibody Protocols: Antibody Labeling Protocols

This protocol describes the covalent coupling of antibodies to biotin. Biotin groups bind with extremely high affinity to streptavidin and avidin, both of which are available commercially coupled with enzymes, fluorescent dyes, or iodine. A biotinylated primary antibody, therefore, can be detected with any of a wide variety of different labels. The biotinylation reaction is simple and mild, and rarely inactivates the antibody. - [Read Labeling Antibodies with Biotin Protocol]

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol details the preparation of biotin-labeled target samples and hybridization of these samples to an Affymetrix in situ synthesized oligonucleotide GeneChip array.  The procedure requires a minimum of 5 µg of purified total RNA as starting material. - [Read Microarray Protocol for Affymetrix In Situ Synthesized Oligo Arrays]