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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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Gel Elongation Assay for Type II Fatty Acid Synthesis Protocol - http://www.natureprotocols.com/2006/06/23/gelelongation_assay_for_type_i.php

Category: Pharmacology and Toxicology Protocols: Drug Discovery

To determine the selectivity of the inhibitors identified during screening efforts we developed gel-elongation assay using crude bacterial lysate directly to determine the target specificities of fatty acid synthesis inhibitors. - [Read Gel Elongation Assay for Type II Fatty Acid Synthesis Protocol]

Gel Mobility Shift Assay Conditions -Mg/EDTA in Gel and Buffer Protocol - http://www.fhcrc.org/science/labs/hahn/methods/biochem_meth/gel_shift.html

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

Protocol for gel mobility shift assay conditions -Mg/EDTA in gel and buffer. - [Read Gel Mobility Shift Assay Conditions -Mg/EDTA in Gel and Buffer Protocol]

Gel Mobility Shift Assay in PDF format - http://www.biochem.northwestern.edu/ibis/morimoto/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/3a.%20GMSA.pdf

Category: DNA Protocols: EMSA DNA-Protein Protocols

This protocol works well for HSF DNA-binding activity experiments. - [Read Gel Mobility Shift Assay in PDF format]

Gel Mobility Shift Assay Protocol - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/3a.%20GMSA.pdf

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

Protocol for gel mobility shift assay. - [Read Gel Mobility Shift Assay Protocol]

Gel Super-Shift Assay EMSA Protocol - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/3b.%20Super-shift.pdf

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

Protocol for gel super shift assay EMSA. - [Read Gel Super-Shift Assay EMSA Protocol]

Gel-elongation Assay for Type II Fatty Acid Synthesis Protocol - http://www.natureprotocols.com/2006/06/23/gelelongation_assay_for_type_i.php

Category: Lipid Protocols

New screening efforts and chemical modifications of existing compounds have been attempted to identify more selective and potent inhibitors. To determine the selectivity of the inhibitors identified during screening efforts we developed gel-elongation assay using crude bacterial lysate directly to determine the target specificities of fatty acid synthesis inhibitors. - [Read Gel-elongation Assay for Type II Fatty Acid Synthesis Protocol]

Gene Expression and Protein Localization in Arabidopsis Protocols - http://www.arabidopsis.org/cshl-course/7-gene_expression.html

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocols for gene expression and protein localization in Arabidopsis. Includes: Detection of the native protein; Detection of a recombinant version; Immunofluorescence detection in Arabidopsis protoplasts; Isolation of Arabidopsis seedling protoplasts; Subcellular localization of GUS-fusion proteins in Arabidopsis seedlings; Localization of Arabidopsis proteins with GUS in situ enzyme assay. - [Read Gene Expression and Protein Localization in Arabidopsis Protocols]

General Method for Ras, Rac, Cdc42, and Rho Activation Assay - http://www.upstate.com/misc/protocol_detail.q.prot.e.general_method_ras.a.name.e.General_Method_for_Ras_Rac_Cdc42_and_Rho_Activation_Assay

Category: Signalling Signal Transduction Protocols

General protocol for Ras, Rac, Cdc42, and Rho activation assay. Includes: Affinity Precipitation/Immunoblot Protocol, Cell Culture and Extract Preparation (Adherent and Non Adherent cells), GTPÎ³S/GDP Loading for Positive and Negative Controls, Ras, Rac ,Cdc42, and Rho Pull-Down Assay and Western Blot and Detection. - [Read General Method for Ras, Rac, Cdc42, and Rho Activation Assay]

Genotyping Protocol: T1320 mouse - http://www.mmrrc.org/strains/425/ctr_protocol.pdf

Category: Molecular Model Organisms: Mouse Animal Protocols: Mouse Genotyping Protocols

Genotyping Protocol: MMRRC 425. Assay Type: PCR - can distinguish heterozygous animals from homozygous animals. DNA Extraction: DNA from tail snips was ... - [Read Genotyping Protocol: T1320 mouse]

Gentle lysis of cells in sucrose-containing Mito-Buffer - http://www.celldeath.de/apometh/mitobuff.html

Category: Cell Biology and Cell Culture: Cell Lysis Protocols

Gentle lysis of mammalian cells for cytochrome c release assay. Based on Yang et al., 1997 Celldeath.de - [Read Gentle lysis of cells in sucrose-containing Mito-Buffer]

Gentle lysis of mammalian cells for cytochrome c release assay - http://www.celldeath.de/apometh/mitobuff.html

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Lysis in the sucrose-containing Buffer ("Mito-Buffer") is supposed to prevent accidential disrupture of the mitochondria to prevent the leakage of mitochondrial proteins (such as cytochrome c) into the cytosol. Celldeath.de - [Read Gentle lysis of mammalian cells for cytochrome c release assay]

GTPÎ³S-induced Inhibition of Binding - http://www.hopkinsmedicine.org/cellbio/devreotes/gtp.htm

Category: Immunology: Chemotaxis

GTPÎ³S-induced Inhibition of Binding. Chemotaxis assay protocols. Devreotes Lab, John Hopkins Medical Centre. - [Read GTPÎ³S-induced Inhibition of Binding]

GUS Reporter Gene Assay Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GUS%20Assays.pdf

Category: Reporter Gene Assays

Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]

Hematopoietic Colony Forming Cell Assays Protocol - http://www.fhcrc.org/science/labs/fero/Protocols/HematopoieticCFC.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the hematopoietic colony forming cell assay. - [Read Hematopoietic Colony Forming Cell Assays Protocol]

High Throughput and Sensitive Assay Measure Yeast Cell Growth Protocol - http://www.natureprotocols.com/2006/11/22/highthroughput_and_sensitive_a.php

Category: Pharmacology and Toxicology Protocols: Drug Discovery

High-throughput and sensitive assay to measure yeast cell growth: a bench protocol for testing genotoxic agents. Method is highly sensitive, provides quantifiable data and offers high-throughput screening capability. Starting with the treatment of cells with different doses of damaging agents, pre-prepared growing media containing 96-well plates are inoculated and cell population is automatically monitored every 10 min for 48 hours. - [Read High Throughput and Sensitive Assay Measure Yeast Cell Growth Protocol]

Histone H1 Kinase Activity Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p910

Category: Signalling Signal Transduction Protocols

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. Includes information: H1 Kinase Assay on Individual Xenopus Oocytes; H1 Kinase Assay on Xenopus Egg Extract Samples; H1 Kinase Assay on Tissue Culture Cells; Helpful protocol hints. - [Read Histone H1 Kinase Activity Assay]

Homogeneous Caspase 3-7 Assay Protocol - http://www.promega.com/tbs/tb295/tb295.pdf

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Homogeneous caspase-3/7 assay. Includes: Detection of Caspase-3/7 Activity in Cell Culture; Detection of Caspase-3 or -7 Activity in Purified Caspase Preparations; Positive and Negative Cell Culture Controls. - [Read Homogeneous Caspase 3-7 Assay Protocol]

How to Assay Light Responses in Neurospora - http://www.fgsc.net/neurosporaprotocols/lightresponses.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to assay light responses in Neurospora. - [Read How to Assay Light Responses in Neurospora]

Human Lymphocyte Cytotoxicity Assay Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/6.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

This method measures the leakage of DNA and lactate dehydrogenase from lymphocytes into the surrounding medium as an indicator of cytotoxicity. This method also includes an assay of intracellular (mitochondrial) diaphorase as a measure of cellular activity (MTT assay). - [Read Human Lymphocyte Cytotoxicity Assay Protocol]

Human T Cell Activation Protocol - http://www.ebioscience.com/ebioscience/appls/AC145.htm#human

Category: Immunology: T Cell Protocols

Protocol for the Stimulation of human peripheral blood mononuclear cells with anti-human CD3 monoclonal antibody; MTT assay for detection of cellular proliferation. Human PBMCs can be activated in vitro by soluble anti-human CD3 antibodies. Performed titration studies with these antibodies and established the following protocol for stimulation of PBMC. - [Read Human T Cell Activation Protocol]

Image Based Assay of Endothelial Cell Tube Formation Protocol - http://www.bdbiosciences.com/pdfs/whitePapers/06-A790030-3A1.pdf

Category: Pharmacology and Toxicology Protocols: Drug Discovery

Protocol describes an assay that relies on the endothelial cells’ ability to form distinct blood-vessel-like tubules in an extracellular matrix where they can subsequently be visualized by fluorescence microscopy. Although quantification of the tubules can be performed by manual tracing, this method precludes the use of the assay in unbiased high-throughput applications. - [Read Image Based Assay of Endothelial Cell Tube Formation Protocol]

Immunodot Assay Protocol - http://wheat.pw.usda.gov/~lazo/methods/goldberg/immuno.html

Category: ELISA Protocols

Protocol for immunodot assay. - [Read Immunodot Assay Protocol]

Immunoprecipitation / Kinase Assay - http://www.upstate.com/misc/protocols.q.prot.e.ipkinaseassay/Immunoprecipitation+%2F+Kinase+Assay

Category: Signalling Signal Transduction Protocols: Protein Kinase Protocols

Immunoprecipitation / Kinase Assay Protocol. Upstate. Protocol applicable only to kinases whose activity is not altered by cell lysis or immunoprecipitation procedures, and do not require soluble cofactors for activity. - [Read Immunoprecipitation / Kinase Assay]

Immunoprecipitation of Endogenous MEKK1 and Kinase Assay Protocol - http://www.natureprotocols.com/2007/01/22/immunoprecipitation_of_endogen.php

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol for immunoprecipitation of endogenous MEKK1 and kinase assay. Includes: Preparation of lysates; Immunoprecipitation; Kinase assay. - [Read Immunoprecipitation of Endogenous MEKK1 and Kinase Assay Protocol]

In Vitro MAP Kinase Assay - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/In%20Vitro%20MAP%20kinase%20assay%20%209.htm

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol describes an in vitro assay to detect MAP kinase activity using PC12 cells. - [Read In Vitro MAP Kinase Assay]

Articles (1-5 of 5)

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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