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ELISA assay - http://www.elisaassay.com

Category: ELISA Protocols

ELISA Assay. Learn about the ELISA assay. Includes ELISA assay protocols and methods. - [Read ELISA assay]

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ChIP Assay Protocol PDF - http://www.bcm.edu/rosenlab/protocols/ChIP.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Formaldehyde cross-linking and chromatin immunoprecipitation assays of tissue culture cells, Based on Boyd and Farnham. Michelle Kallesen, Rosen Lab. - [Read ChIP Assay Protocol PDF]

ChIP on ChIP Protocol PDF - http://genomecenter.ucdavis.edu/farnham/farnham/protocols/MOJO%20Dec%202003%20ChIP-chip%20protocol.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Method to identify genomic targets of DNA-binding factors. Chromatin immunoprecipitation (ChIP) assay on high-throughput microarray based methods for discovering genomic regions occupied by human DNA-binding proteins. Oberley and Farnham. - [Read ChIP on ChIP Protocol PDF]

ChIP ping for Dummies - http://mcardle.oncology.wisc.edu/sugden/Protocols/html%20files/ChIPping%20for%20Dummies.htm

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Very nice and detailed ChIP assay protocol procedure. Blocking Staph A cells, Crosslinking cells, Chromatin preparation from cell pellets, sonication, blocking. Sugden Laboratory - [Read ChIP ping for Dummies]

Chromatin Immunoprecipitation (ChIP) Assay - http://www.upstate.com/misc/protocol_detail.q.prot.e.chrom_ip.a.name.e.Chromatin_Immunoprecipitation_Assay

Category: Epigenetics and Methylation Protocols: ChIP Chromatin Immunoprecipitation Protocols

Chromatin Immunoprecipitation (ChIP) Assay protocol. Millipore. Includes protocols on Optimization of DNA Shearing, Sonication and PCR for ChIP. - [Read Chromatin Immunoprecipitation (ChIP) Assay]

Chromatin Immunoprecipitation (ChIPs) Protocol - http://genomecenter.ucdavis.edu/farnham/farnham/protocols/chips.html

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

IP Wash buffer recipes and protocol for ChIP Assay. Protocol works with NIH 3T3, Friend, HeLa, Raji and CHO cells.. Farnham Lab - [Read Chromatin Immunoprecipitation (ChIPs) Protocol]

Chromatin IP (CHIP assay) - http://www.fhcrc.org/science/labs/breeden/Methods/chromatinIP.html

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

ChIP assay protocol with 2 steps: in vivo formaldehyde cross-linking of whole cells and protein-protein and protein-DNA interactions, followed by immunoprecipitation of protein-DNA complexes with specific antibodies from sonicated extracts. Breeden Lab - [Read Chromatin IP (CHIP assay)]

Clonality - X Chromosome Inactivation Assay Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/DNA/DNAProtocols/Clonality.html

Category: Genetics and Genomics: Cancer Genetics Protocols

Investigators can utilize X chromosome inactivation (methylation) to determine the clonality status of a tumor or premalignant lesion in females. The technique is based on a methylation-sensitive restriction enzyme and analysis of a polymorphic locus on the X chromosome. Clonal cell populations will show "loss" of the non-methylated allele after restriction digest. The assay can be performed on DNA recovered from microdissected samples. Both frozen tissue and fixed-embedded tissue can be used. - [Read Clonality - X Chromosome Inactivation Assay Protocol]

Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes - http://76.162.25.80/bpo/arts/1/48/m48.pdf

Category: E Coli Protocols: E coli Genetics Protocols

Protocol for Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase. - [Read Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes]

Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4342

Category: Reporter Gene Assays: Luciferase Assay Protocols

It is possible that some cell lines have lost the ability to perform RNAi or that cells derived from certain tissues do not support RNAi. This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4342

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Cytochrome P450 Assay Protocol - http://www.promega.com/tbs/tb325/tb325.pdf

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol for cytochrome P450 assay. - [Read Cytochrome P450 Assay Protocol]

Cytochrome P450 CYP450 Assay Protocol - http://www.promega.com/tbs/ep026/ep026.pdf

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol for cytochrome P450 CYP450 assay. - [Read Cytochrome P450 CYP450 Assay Protocol]

CytoTox-ONE Homogeneous Membrane Integrity Assay Protocol - http://www.promega.com/tbs/ep016/ep016.pdf

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Protocol describes automation of the CytoTox-ONE Homogeneous Membrane Integrity Assay. - [Read CytoTox-ONE Homogeneous Membrane Integrity Assay Protocol]

Deadenylation Assay Protocol - http://www.cvmbs.colostate.edu/mip/wiluszlab/protocols/deadenylase.htm

Category: Nucleic Acid Modification Protocols: Nucleic Acid Radiolabeling Protocols

Protocol for deadenylation assay. - [Read Deadenylation Assay Protocol]

Detailed NF-kB EMSA Protocol - http://www.celldeath.de/apometh/emsa.html

Category: Cell Biology and Cell Culture: Apoptosis Protocols: NF-kB EMSA Protocols

Detailed NF-kB EMSA Protocol. Includes Nuclear extract preparation, Labeling of the oligonucleotide probe: T4 Polynucleotide Kinase Reaction, Supershift assay to identify NF-kB, and recipes for EMSA buffers. Celldeath.de - [Read Detailed NF-kB EMSA Protocol]

Detection of Protein Phosphorylation in Tissues and Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&objectId=6677CD53C088A0E7A7B22F422FFCDC12&treeNodeId=9E6639B7FED0C828BBFE860A8E957694

Category: Immunology: Immunoprecipitation Protocols: Immunoprecipitation of Radiolabeled Cells Protocols

Most powerful and convincing method to determine if a specific protein is phosphorylated in a physiologically relevant manner is to assay phosphorylation in situ. The procedure described involves incubating cultured cells (e.g., primary neuronal cultures or transfected cells) or tissue preparations (e.g., hippocampal slices) with [32P]orthophosphate, which is then taken up by the cells or tissues and incorporated into the γ-phosphate position of ATP. - [Read Detection of Protein Phosphorylation in Tissues and Cells Protocol]

Detection of Recombinant Proteins by ELISA assay - http://omrf.ouhsc.edu/~frank/ELISA.html

Category: ELISA Protocols

Detection of Recombinant Proteins by ELISA protocol. Dr. Frank, Oklahoma Medical Research Foundation. - [Read Detection of Recombinant Proteins by ELISA assay]

Detection of siRNA in C. elegans Using Rnase Protection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4319

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol uses Rnase protection to detect short interfering RNAs (siRNAs) in RNA preparations from Caenorhabditis elegans. SiRNAs can also be detected by northern blot. However, the Rnase protection assay seems to be more sensitive. - [Read Detection of siRNA in C. elegans Using Rnase Protection Protocol]

Detection of Ubiquitylated Proteins in Yeast Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4615

Category: Yeast Protocols: Yeast Protein Protocols

This assay is performed to detect ubiquitylated proteins in yeast. Yeast that have been transformed with a vector expressing polyhistidine-tagged ubiquitin (Ub) under the control of a copper-inducible promoter are grown, induced with copper, and harvested. Total ubiquitylated proteins are then recovered by nickel-affinity chromatography, and specific proteins are detected by Western blotting. - [Read Detection of Ubiquitylated Proteins in Yeast Protocol]

Determination of Chlorophyll Content in Chloroplasts - http://www.bio.com/protocolstools/protocol.jhtml?id=p1285

Category: Plant Biology Protocols: Photosynthesis and Respiration Protocols

This protocol describes how to determine the chlorophyll content of chloroplasts using a spectrophotometric assay. - [Read Determination of Chlorophyll Content in Chloroplasts]

Determination of the CpG Island Methylator Phenotype (CIMP) in Colorectal Cancer using MethyLight - http://www.natureprotocols.com/2006/06/30/determination_of_the_cpg_islan.php

Category: Genetics and Genomics: Cancer Genetics Protocols

Detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in Colorectal Cancer using MethyLight]

Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight - http://www.natureprotocols.com/2006/06/30/determination_of_the_cpg_islan.php

Category: Nucleic Acid Modification Protocols: DNA Methylation Protocols

Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight. Protocol describes a detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight]

Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight - http://www.natureprotocols.com/2006/06/30/determination_of_the_cpg_islan.php

Category: PCR Protocols: Real-Time PCR Protocols

Protocol provides a detailed, step-by-step protocol of the MethyLight assay for detection of CIMP with high sensitivity and specificity in colorectal cancer using a five marker panel composed of CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. - [Read Determination of the CpG Island Methylator Phenotype (CIMP) in colorectal cancer using MethyLight]

Determining Number of Live and Dead Cells in Cell Population: Cytotoxicity Assay Protocol - http://www.promega.com/paguide/chap4.htm#title4

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Cell-based assays are important tools for contemporary biology and drug discovery because of their predictive potential for in vivo applications.This assay gives ratiometric, inversely proportional values of viability and cytotoxicity (Figure 4.15) that are useful for normalizing data to cell number. Also, this reagent is compatible with additional fluorescent and luminescent chemistries. - [Read Determining Number of Live and Dead Cells in Cell Population: Cytotoxicity Assay Protocol]

DNA laddering assay for treated cells - http://www.celldeath.de/apometh/dnaladd.html

Category: Cell Biology and Cell Culture: Apoptosis Protocols

DNA laddering can be detected from samples with only 8% apoptotic cells. Alternatively, the cells can be stained with DAPI and analysed by flow cytometry. CellDeath.de - [Read DNA laddering assay for treated cells]

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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.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

Histone H1 Kinase Activity Assay

Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.

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