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Antigen Retrieval - Immunohistochemistry - http://axon.med.harvard.edu/~cepko/protocol/mike/A2.html

Category: Immunology: Antigen Retrieval

Denaturing Solution and Procedure for AR. Mike Dyer. - [Read Antigen Retrieval - Immunohistochemistry]

Denaturing Agarose Gel Electrophoresis of RNA - http://www.ambion.com/techlib/append/supp/rna_gel.html

Category: Molecular Biology Protocols: Agarose Gel Preparation

The quality of an RNA preparation may be assessed by electrophoresis on a denaturing agarose gel. - [Read Denaturing Agarose Gel Electrophoresis of RNA]

Denaturing Agarose Gel Electrophoresis of RNA Protocol - http://www.ambion.com/techlib/append/supp/rna_gel.html

Category: Nucleic Acid Electrophoresis Protocols: RNA Electrophoresis Protocols

Protocol for denaturing agarose gel electrophoresis of RNA. Includes: Prepare the gel; Prepare the RNA sample; Electrophoresis. - [Read Denaturing Agarose Gel Electrophoresis of RNA Protocol]

Denaturing Protein Immunoprecipitation from Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4619

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes a denaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. The use of denaturing IPs reduces background considerably. - [Read Denaturing Protein Immunoprecipitation from Mammalian Cells Protocol]

Denaturing Protein Immunoprecipitation from Yeast Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4643

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Immunoprecipitation Protocols

The use of denaturing Ips reduces background and simplifies protein harvest. Yeast are boiled in an SDS-containing solution to liberate total proteins. - [Read Denaturing Protein Immunoprecipitation from Yeast Protocol]

High-resolution SNP mapping by denaturing HPLC - http://www.pnas.org/cgi/content/full/99/16/10575

Category: DNA Protocols: SNP Protocols

High-resolution SNP mapping by denaturing HPLC. A SNP mapping procedure that relies on resolving polymorphisms by denaturing HPLC without the necessity of determining the nature of the SNPs. They demonstrate the use of denaturing high-performance liquid chromatography to identify mutations in the candidate genes and to fine-map chromosomal breakpoints. - [Read High-resolution SNP mapping by denaturing HPLC]

Immunoprecipitation with Anti-Phosphotyrosine:Biotin Conjugates Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Biotin_Conjugates.shtml

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]

Immunoprecipitation with Antibody:Agarose Conjugates Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Agarose_Conjugates.shtml

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with antibody:agarose conjugates. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation with Antibody:Agarose Conjugates Protocol]

Immunoprecipitation With Soluble Antibodies Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Soluble_Antibodies.shtml

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with soluble antibodies. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation With Soluble Antibodies Protocol]

Immunoprecipitation: Denaturing Lysis Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4534

Category: Immunology: Immunoprecipitation Protocols

For many sources of antigens, one useful method of lysis is to treat cells with harsh, denaturing solutions to release most of the protein antigens, as described here. The lysates are then diluted to reduce the denaturing conditions to levels that are suitable for the formation of antibody-antigen complexes. The resulting solution is precleared prior to immunoprecipitation. - [Read Immunoprecipitation: Denaturing Lysis Protocol]

Non-denaturing Cell Lysis Buffer - http://www.chemicon.com/techsupp/celllysis.asp

Category: Buffers Media, and Solutions: Buffer Recipes for Protein Work

Non-denaturing Cell Lysis Buffer. Chemicon. - [Read Non-denaturing Cell Lysis Buffer]

Nondenaturing Protein Immunoprecipitation from Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4620

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes a nondenaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. However, the nondenaturing protocol is useful when one wishes to separate soluble from insoluble proteins, or when the antibody being used recognizes a native epitope. - [Read Nondenaturing Protein Immunoprecipitation from Mammalian Cells Protocol]

PCR primer design and pcr reaction optimization. - http://www.mcb.uct.ac.za/pcroptim.htm

Category: PCR Protocols: PCR Optimization

PCR primer design and pcr reaction optimization. Ed Rybicki. Factors Affecting the PCR, Denaturing Temperature and Time, Annealing Temperature and Primer Design, Primer Length, Degenerate Primers, Elongation , Temperature and Time, Reaction Buffer, Cycl - [Read PCR primer design and pcr reaction optimization.]

Preparation of a Denaturing Agarose Gel for the Analysis of RNA Samples Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4127

Category: Nucleic Acid Electrophoresis Protocols: RNA Electrophoresis Protocols

Protocol for preparation of a denaturing agarose gel for the analysis of RNA samples. The integrity of RNA samples can be analyzed by electrophoresis on a denaturing agarose gel containing formaldehyde. - [Read Preparation of a Denaturing Agarose Gel for the Analysis of RNA Samples Protocol]

Preparation of Denaturing Polyacrylamide Gels Containing Formamide Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3807

Category: DNA Protocols: DNA Sequencing Protocols

The inclusion of formamide in sequencing gels eliminates secondarystructure in the DNA during electrophoresis. Formamide gelsare particularly useful and almost a necessity when sequencingDNA templates with a G/C content >55%. - [Read Preparation of Denaturing Polyacrylamide Gels Containing Formamide Protocol]

Preparation of Denaturing Polyacrylamide Gels Containing Formamide Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3807

Category: Nucleic Acid Electrophoresis Protocols

Protocol for preparation of denaturing polyacrylamide gels containing formamide.The inclusion of formamide in sequencing gels eliminates secondary structure in the DNA during electrophoresis. Formamide gels are particularly useful and almost a necessity when sequencing DNA templates with a G/C content >55%. - [Read Preparation of Denaturing Polyacrylamide Gels Containing Formamide Protocol]

Preparation of Denaturing Polyacrylamide Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3793

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Polyacrylamide Gels Protocols

Protocol for preparation of denaturing polyacrylamide gels. - [Read Preparation of Denaturing Polyacrylamide Gels Protocol]

Protocol for Immunopurification of Tyrosine Phosphorylated Proteins - http://www.bdbiosciences.com/pharmingen/protocols/Tyrosine_Phosphorylated_Proteins.shtml

Category: Signalling Signal Transduction Protocols

Protocol for immunopurification of tyrosine phosphorylated proteins. Protocol includes: Cell Lysis (Non-Denaturing and Denaturing Conditions) and Purification. - [Read Protocol for Immunopurification of Tyrosine Phosphorylated Proteins]

Pulse-Chase Assay for Measuring Protein Stability in Yeast Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4641

Category: Yeast Protocols: Yeast Protein Protocols

This assay directly measures the stability of a specific protein in yeast. Following pulse-labeling of yeast cells, the target protein is recovered by denaturing immunoprecipitation. - [Read Pulse-Chase Assay for Measuring Protein Stability in Yeast Protocol]

Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4221

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Immobilized metal-ion affinity chromatography (IMAC) is suitable for the purification of proteins under denaturing conditions. Either guanidine-HCl or urea can be used, although guanidine-HCl is a stronger denaturant than urea. Proteins that have been adsorbed to the column in the presence of guanidine-binding buffer may be washed with urea-binding buffer and eluted with urea elution buffer. - [Read Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol]

Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4109

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles. Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number). Total RNA is used as an internal control. - [Read Relative RT-PCR: Determining the Linear Range of Amplification and Optimizing the Primers:Competimer]

Single-Strand Conformation Polymorphism Analysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4118

Category: Genetics and Genomics: Cancer Genetics Protocols: Single Strand Conformational Polymorphism(SSCP) Protocols

Single-strand confirmation polymorphism analysis (SSCP) is a powerful and robust method for the detection of DNA sequence changes (single-base substitutions) based on shifts in electrophoretic mobility. In this protocol, the target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. - [Read Single-Strand Conformation Polymorphism Analysis Protocol]

Single-Strand Conformation Polymorphism Analysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4118

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

The target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. Differences in sequence alter the conformation of the DNA and hence its electrophoretic mobility and, because of the high resolution of polyacrylamide gels, most conformational changes caused by subtle changes in sequence can be detected. - [Read Single-Strand Conformation Polymorphism Analysis Protocol]

Storage of Sera Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4279

Category: Antibody Protocols: Antibody Handling Protocols

This protocol describes the methods of storage for antibody-containing sera. Antibodies are resistant to a broad range of mildly denaturing conditions, so long-term storage is relatively easy. - [Read Storage of Sera Protocol]

Synthesis of Single-stranded DNA Probes of Heterogeneous Length from Bacteriophage M13 Templates - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3546

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Technique yields a heterogeneous population of short radiolabeled molecules 200-300 nucleotides in length. These probes are synthesized, as in Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates, by extension of an oligonucleotide primer on a single-stranded DNA template. The radiolabeled products of the reaction are then separated from the template by electrophoresis through a denaturing gel from which they are eluted directly into hybridization buffer. - [Read Synthesis of Single-stranded DNA Probes of Heterogeneous Length from Bacteriophage M13 Templates]

Articles (1-2 of 2)

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.