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96-Well Confirmation Yeast PCR Protocol - http://sequence-www.stanford.edu/group/yeast_deletion_project/96_well_confirmation.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for 96-well confirmation Yeast PCR. Includes: Clonal purification; Generate a master plate (96-well format); Making a frozen backup stock; Confirmation PCR for one Row; ORF Specific Confirmation PCR --> "A-B" primers (upstream junction); Transfer template DNA to multiwell PCR plate; Prepare and dispense master mix for A-B PCR. - [Read 96-Well Confirmation Yeast PCR Protocol]

A Self-generated Gradient to Discriminate a Cytosolic or Membrane Location of Large Protein Complex - http://www.axis-shield.com/densityhome/optiprep/S24.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Protocol describes systems in which the membranes are allowed to float through a density gradient from a dense load zone. This is an ideal format. - [Read A Self-generated Gradient to Discriminate a Cytosolic or Membrane Location of Large Protein Complex]

Analysis of Proteins using Small Format 2D Gel Electrophoresis - http://www.abdn.ac.uk/~mmb023/protocol.htm

Category: Proteomic Protocols: 2-D Gel Electrophoresis

Analysis of Proteins using Small Format 2D Gel Electrophoresis. Cash Lab - [Read Analysis of Proteins using Small Format 2D Gel Electrophoresis]

Assay of Intracellular Free Calcium in RAW 264.7 Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000210.pdf?pid=PP00000210

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i, in cultured adherent RAW 264.7 cells, using a 96-well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fluo-3, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells]

Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fluo-3 Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000210.pdf?pid=PP00000210

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i, in cultured adherent RAW 264.7 cells, using a 96- well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fluo-3, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. Fluorescence for the adherent cells is measured over time by using a bottom read of a 96-well plate, with cells that have been washed. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fluo-3 Protocol]

Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 (with FLEXstation) - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000211.pdf?pid=PP00000211

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+], in cultured adherent RAW 264.7 cells, using a 96-well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fura-2, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 (with FLEXstation)]

Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000211.pdf?pid=PP00000211

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i , in cultured adherent RAW 264.7 cells, using a 96- well plate format. This objective is accomplished by using the Ca2+-sensitive fluorescent dye, fura-2, which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. - [Read Assay of Intracellular Free Calcium in RAW 264.7 Cells Loaded with Fura-2 Protocol]

Comparison Different Protein Quantitation Determination Methods - http://wolfson.huji.ac.il/purification/Protocols/Comparision_of_Methods.htm

Category: Protein Protocols: Protein Quantitation Concentration Protocols

A Comparision of Different Protein Determination Methods and protocols in table format. The Wolfson Centre for Applied Structural Biology The Hebrew University of Jerusalem Dr. Mario Lebendiker. - [Read Comparison Different Protein Quantitation Determination Methods]

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]

MAKING MICROARRAY PRINTING PLATES (IN DMSO) - http://pga.tigr.org/sop/M003_1a.pdf

Category: DNA Microarray Protocols

This protocol describes the method for making microarray printing plates in a 96 well format. (The same procedure applies to a 384 well format as well.) Hasseman. TIGR Microarray Protocols - [Read MAKING MICROARRAY PRINTING PLATES (IN DMSO)]

Measurement of Cell Adhesion Under Static Conditions - http://www.glycotech.com/protocols/Proto8.html

Category: Cell Biology and Cell Culture: Cell Adhesion Protocols

Many proteins and molecules promote cell adhesion including several cell surface carbohydrate binding proteins. Cell adhesion measurements on 96-well microtiter plate format are difficult due to the shear forces generated by washing the wells. The protocol here introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity. This eliminates uncontrolled shear forces and passage of adherent cells through a liquid/air interface. John L. Magnani~GlycoTech Corporation. - [Read Measurement of Cell Adhesion Under Static Conditions]

MICROARRAY cDNA CLONE GROWTH AND TEMPLATE - http://pga.tigr.org/sop/M001_1a.pdf

Category: DNA Microarray Protocols

This protocol describes clone handling, plate replication, and DNA template preparation in a 96 well format. Hasseman. TIGR Microarray Protocols - [Read MICROARRAY cDNA CLONE GROWTH AND TEMPLATE]

RNAi Screens in C. elegans Protocol - http://www.natureprotocols.com/2006/07/19/rnai_screens_in_c_elegans_in_a.php

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol for RNAi screens in C. elegans in a 96-well liquid format and their application to the systematic identification of genetic interactions. The procedure allows thousands of RNAi feeding experiments to be performed per investigator per day. - [Read RNAi Screens in C. elegans Protocol]

Use of the Bradford Protein Assay in a Microtiter Plate Format - http://www.animal.ufl.edu/hansen/protocols/minibradford.htm

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

Use of the Bradford Protein Assay in a Microtiter Plate Format Protocol. Includes a short introduction to the Bradford assy, materials and procedure. - [Read Use of the Bradford Protein Assay in a Microtiter Plate Format]

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.

Quick DNA Agarose Gel Electrophoresis Protocol.

A quick and simple protocol for the electrophoresis of DNA fragments using agarose gels.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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