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2-D Polyacrylamide Gel Electrophoresis Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/2DPage.html

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

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

2x Lysis Buffer Recipe - http://utzlab.stanford.edu/publications/lysisbuffer.doc

Category: Proteomic Protocols: 2-D Gel Electrophoresis

2x Lysis Buffer Recipe. Utz Lab. - [Read 2x Lysis Buffer Recipe]

Acetone precipitation of protein from Buffer RLT or Buffer RLT - http://www1.qiagen.com/literature/protocols/pdf/RY22.pdf

Category: Protein Protocols: Protein Precipitation Procols

Acetone precipitation of protein from Buffer RLT or Buffer RLT. QIAGEN. - [Read Acetone precipitation of protein from Buffer RLT or Buffer RLT]

Activation of p70s6k and MAP Kinases: IP and Kinase Activity - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/ACTIVATION%20OF%20p70s6k%20and%20MAP%20KINASES.htm

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol describes the activation of MAP Kinase activity. Protocol contains information on: Buffers: ST, STE, Lysis Buffer Buffer A, Buffer B and 10x Kinase Buffer. - [Read Activation of p70s6k and MAP Kinases: IP and Kinase Activity]

Akt/PKB Kinase Assay - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/Akt%20PKB%20kinase%20assay.htm

Category: Signalling Signal Transduction Protocols

Protocol for Akt/PKB kinase assay. Protocol includes: Akt lysis buffer; Kinase Buffer; 1.5x Akt kinase reaction buffer; Start solution; Lipofectamine PLUS; CDNA synthesis (Roche #1483?88). - [Read Akt/PKB Kinase Assay]

Alkaline Phosphatase (APAAP) Technique - http://www.nottingham.ac.uk/pathology/protocols/apap.html

Category: Signalling Signal Transduction Protocols

Technique for Alkaline Phosphatase (APAAP). Techniques includes information on: Cytological Preparations, Fixation, Technique, Results, Substrate Solution and 0.005M Tris/HC1 Saline Buffer pH 7.6. - [Read Alkaline Phosphatase (APAAP) Technique]

Analyzing Yeast Colonies by PCR Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4015

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

Yeast colonies are suspended in complete PCR buffer and transferred to a thermal cycler for 35 cycles of PCR. The products of the amplification reaction are analyzed by gel electrophoresis. - [Read Analyzing Yeast Colonies by PCR Protocol]

Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography - http://www.upstate.com/misc/protocol_detail.q.prot.e.antibody_purification.a.name.e.Antibody_Purification

Category: Protein Protocols: Antibody Purification Protocols

Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Buffer Preparation, Preparation of a Protein A Agarose or Protein G Agarose Affinity Column, Pouring the Protein A/G Affinity Column, Preparation of Antiserum or Ascites for Affinity Chromatography, Affinity Chromatography Using Protein A/G Agarose. - [Read Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography]

Assay for Luciferase in Extracts of Mammalian Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3642

Category: Reporter Gene Assays: Luciferase Assay Protocols

In this protocol, cells transfected with a luciferase reporter plasmid are lysed in a detergent-containing buffer. Luciferase in the extract catalyzes an oxidation reaction in which D-luciferin is converted to oxyluciferin, with production of light at 556 nm that can be quantified in a luminometer. - [Read Assay for Luciferase in Extracts of Mammalian Cells Protocol]

B-GAL Filter Assay Protocol - http://www.fhcrc.org/science/labs/breeden/Methods/b-GAL_filterassay.html

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

Protocol for B-Gal filter assay. Includes Z buffer and X-Gal solution. - [Read B-GAL Filter Assay Protocol]

Buffer calculator tool - http://molbiol.ru/eng/protocol/01_07.html

Category: Buffers Media, and Solutions

Buffer calculator tool for commonly used buffer compound solutions. Making different volumes? use this! (Practical Molecular Biology) - [Read Buffer calculator tool]

Buffer Calculator. Lab Velocity - http://researchlink.labvelocity.com/tools/bufferCalculator.jhtml

Category: Buffers Media, and Solutions

Buffer Calculator. Amazing calculator input whatever you want. Time saver! Lab Velocity. - [Read Buffer Calculator. Lab Velocity]

Buffer Recipies for 2D-Electrophoresis - http://www.queens-pfd.ca/images/pdfs/2DE%20outline.pdf

Category: Buffers Media, and Solutions

Buffer Recipies for 2D-Electrophoresis - [Read Buffer Recipies for 2D-Electrophoresis]

Buffer Solutions and How they work... - http://www.chemguide.co.uk/physical/acidbaseeqia/buffers.html

Category: Buffers Media, and Solutions: Buffer Theory

This page describes simple acidic and alkaline buffer solutions and explains how they work. - [Read Buffer Solutions and How they work...]

CaspaTag Caspase Activity Protocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/caspatag.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

This assay is based on carboxyfluorescein labeled fluoromethyl ketone (FMK)-peptide inhibitors of caspases. Includes: Working Dilution of FMK-peptide inhibitors; 1X Working Dilution Wash Buffer; Protocol for Flow Cytometry. - [Read CaspaTag Caspase Activity Protocol]

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]

Common Buffer List - http://www.bio.unc.edu/faculty/salmon/lab/protocolscommonbuffers.html

Category: Buffers Media, and Solutions

Salmon Lab, University of North Carolina at Chapel Hill, Department, of Biology. - [Read Common Buffer List]

Common Buffers - http://wheat.pw.usda.gov/~lazo/methods/lazo/buffer.html

Category: Buffers Media, and Solutions

Recipes for commonly used buffer solutions. (Gerard R. Lazo) - [Read Common Buffers]

DNA Affinity Chromatography Using Gravity Flow - Subscription Required - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4206

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

DNA Affinity Chromatography, DNA affinity chromatography can be a low-tech method using gravity flow at 4Â°C, a disposable chromatography column, and DNA affinity resin prepared in the laboratory (see Preparation of a DNA Affinity Column). Include 10-20% glycerol and 0.025-0.1% NP-40 in the column buffers to suppress losses due to nonspecific adsorption of protein to surfaces. Load the protein in a buffer that is compatible with binding of the protein to its target site. Keith Brocklehurst et al - [Read DNA Affinity Chromatography Using Gravity Flow - Subscription Required]

DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4098

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

In this protocol, the DNA-binding capacity of Wizard MagneSil particles is used to capture and release a consistent amount of DNA (100 ng) across a wide range of samples. At the end of the procedure, the DNA is eluted into 100 µl Elution Buffer to give a final concentration of 1 ng/µl, relieving the need for postpurification DNA quantitation. - [Read DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol]

DNA ligation with Linearized DNA - http://www.genome.ou.edu/protocol_book/protocol_partII.html#II.D

Category: Molecular Biology Protocols: DNA Ligation Protocols

DNA ligations are performed by incubating DNA insert with linearized cloning vector in the presence of ligation buffer, rATP, and T4 DNA ligase. Roe. Univ. Oklahoma. - [Read DNA ligation with Linearized DNA]

DNA Preparation from Blood PDF - http://www.riedlab.nci.nih.gov/publications/2438.2413_DNA_Prep_Blood.pdf

Category: DNA Protocols: DNA Extraction Blood

Method for DNA Preparation from Blood. Using lysis buffer and phenol/chloroform/isoamyl alcohol. Thomas Ried, NCI. - [Read DNA Preparation from Blood PDF]

Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization Protocol - https://commerce.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeId=3DD21299A00419DEBAFDBC78E47CBE79

Category: PCR Protocols: PCR Optimization

Method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield. The first step of PCR simply entails mixing template DNA, two appropriate oligonucleotide primers, Taq or other thermostable DNA polymerases, deoxyribonucleoside triphosphates (dNTPs), and a buffer. - [Read Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization Protocol]

ENZYME-ASSISTED IMMUNOELECTROBLOTTING (IEB OR WESTERN BLOTTING) - http://www.mcb.uct.ac.za/western.htm

Category: Protein Protocols: Western Blot Protocol

ENZYME-ASSISTED IMMUNOELECTROBLOTTING (IEB OR WESTERN BLOTTING. E.P. Rybicki and Maud Purves. Department of Microbiology. Copper Staining of Gels, Indirect enzyme immunoassay, Staining Proteins on Membranes, Blotting Buffer, Staining of proteins in gels - [Read ENZYME-ASSISTED IMMUNOELECTROBLOTTING (IEB OR WESTERN BLOTTING)]

Extraction and Solubilization of Total Protein from Microorganisms Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4224

Category: Bacterial Cell Culture Protocols

The growth conditions of microbial cell cultures and the time of sample collection should be optimized and standardized when growing cells for protein extraction. Because cells may excrete proteases and other extracellular enzymes, and compounds in the medium may interfere with extraction, wash cultures with an isotonic buffer, such as PBS or sucrose before solubilization. - [Read Extraction and Solubilization of Total Protein from Microorganisms Protocol]

Articles (1-10 of 23)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

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.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

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.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

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.

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