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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]

A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb16/16183-2.pdf

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The protocol includes: organelle isolation, deoxyribonuclease treatment, lysis, deproteinisation and a final DNA purification with sodium dodecyl sulphate and potassium acetate. The organelle DNA yield is 5–10 micrograms per gram of tissue and the DNA is fully restrictable. The technique is inexpensive and appropriate for the isolation of multiple samples of organelle DNA from a small amount of tissue. - [Read A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower]

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 Lysis Mini-Prep Protocol - http://preuss.bsd.uchicago.edu/protocols/Alkaline.html

Category: Microbiology: Mini-Prep Plasmid Purification

Harvesting and Lysis of Bacteria. Mini-prep plasmid purification using Solution I, Solution II and Solution III. The Preuss Lab. Univ. Chicago. - [Read Alkaline Lysis Mini-Prep Protocol]

Bacterial Media Solutions and Stocks - http://humgen.wustl.edu/hdk_lab_manual/plasmid/plsmid11.html

Category: Microbiology: Bacterial E.Coli Culture Media & Plates

agar, Ampicillin Stock, Alkaline Lysis Solution, SOB, SOC media, and X-gal stock solutions. Donis Keller. - [Read Bacterial Media Solutions and Stocks]

Baculovirally Infected Sf9 Cell Lysis Protocol - http://ruummc.med.uu.nl/subsite/RepHTML/Protocols/Sf9lysis.html

Category: Cloning Protocols: Baculovirus Protein Expression Protocols

Protocol for Baculovirally infected Sf9 cell lysis. Includes: Harvesting; Lysis. - [Read Baculovirally Infected Sf9 Cell Lysis Protocol]

Cell Lysate Extracts - http://www.upstate.com/misc/protocol_detail.q.prot.e.CellLysate_Prep.a.name.e.Cell_Lysate_Extracts-General_Protocols

Category: Protein Protocols: Protein Extraction From Cells

Cell Lysate Extracts. Great protocols for cell lysis preparation from a variety of cell types. There are numerous methods of cell stimulation and lysis. For a given protein, Upstateâ€™s Laboratories determine the specific treatment upon initial testing of its products. It is important to select the correct cell line, stimulation procedure (if any), and lysis protocol. Upstate. - [Read Cell Lysate Extracts]

CHO Centrosome Prep Protocol - http://mitchison.med.harvard.edu/protocols/mt1.html

Category: Cell Organelle Protocols: Nucleus Protocols

The key step is the lysis which solubilizes centrosomes away from nuclei by very low ionic strength lysis after treatment of cells with nocodazole and cytochalasin B. The released centrosomes are then centrifuged onto a Ficoll cushion (to avoid pelleting) and the interface between the lysate and the Ficoll is collected and the centrosomes are concentrated on a sucrose gradient. Fractions are assayed by spindown and double IF with 5051 serum and anti-tubulin and the pooled fractions are frozen... - [Read CHO Centrosome Prep Protocol]

Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol - http://www.nature.com/nprot/journal/v1/n2/abs/nprot.2006.98.html

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. - [Read Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol]

DNA Isolation From BAC & PAC Clones Protocol - http://bacpac.chori.org/bacpacmini.htm

Category: BACs Bacterial Artificial Chromosome Protocols

Rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

DNA Isolation From BAC & PAC Clones Protocol - http://bacpac.chori.org/bacpacmini.htm

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

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]

Dual Luciferase Reporter System Protocol (PROMEGA) - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/II.%20Eukaryotes/F.%20Luciferase%20Assays/2b.Dual%20luciferase.pdf

Category: Reporter Gene Assays: Luciferase Assay Protocols

Protocol for dual luciferase reporter system. Includes: Transfections; Cell Lysis; Reagent Preparation; Luminescence measurement; Wash injectors. - [Read Dual Luciferase Reporter System Protocol (PROMEGA)]

Extraction of Geminiviral DNA from a Highly Mucilaginous Plant Protocol - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb18/R00-056.pdf

Category: Viral Protocols

Protocol for the extraction of geminiviral DNA from a highly mucilaginous plant (Abelmoschus esculentus). Includes: Extraction; Alkali lysis; Cloning. - [Read Extraction of Geminiviral DNA from a Highly Mucilaginous Plant Protocol]

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]

GST Fusion Protein Purification from Yeast - http://www.med.unc.edu/~hdohlman/gst.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

GST Fusion Protein Purification from Yeast. Lysis Buffer recipe for yeast, glutathione bead method for GST purification. Dohlman Lab - [Read GST Fusion Protein Purification from Yeast]

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: 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]

Induction and Measurement of Cytotoxic T Lymphocyte Activity Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663458DAAA03E5A23DC69D6BE8BE3E&objectid=6674B15EE255310CC4C2307B31AE3411

Category: Immunology: T Cell Protocols

Describes generating CTL against some commonly used target antigens. Two methods for the quantitation of CTL activity are described based on the two pathways used bt CTL to kill target cells. In one pathway, they release lytic granules containing perforin and granzymes, leading to apoptosis and target cell lysis. In a second pathway, they trigger apoptosis via Fas/Fas ligand interactions. - [Read Induction and Measurement of Cytotoxic T Lymphocyte Activity Protocol]

Isolation of BAC DNA from Small-scale Cultures Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4006

Category: BACs Bacterial Artificial Chromosome Protocols

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA. - [Read Isolation of BAC DNA from Small-scale Cultures Protocol]

Lambda plate lysis Protocol - http://www.rpgroup.caltech.edu/~paul/protocols/phage/lambda/plate/

Category: Molecular Biology Protocols: Phage Protocols and Methods: Lambda Phage Protocols

Phage Protocols - Lambda plate lysis. Rob Philip Group. - [Read Lambda plate lysis Protocol]

Luciferase Assay 24 Well Plate (Promega System) - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/II.%20Eukaryotes/F.%20Luciferase%20Assays/2a.single%20luciferase.pdf

Category: Reporter Gene Assays: Luciferase Assay Protocols

Luciferase assay using a 24 well plate. Includes: Cell Lysis; Reagent Preparation; Luminescence measurement; Wash injector; Protein quantitation; Data Analysis. - [Read Luciferase Assay 24 Well Plate (Promega System)]

Articles (1-10 of 11)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Lambda Phage Protocol for Large Preparation DNA Recovery

Lambda Phage Protocol for Large Preparations DNA Recovery.

Lambda Phage Protocol Large DNA Preparation

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.

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.

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

Toluidine Blue Stain

Toluidine Blue staining method and protocol for cell or tissue histology.

Thionin Stain

Thionin staining method and protocol for cell or tissue histology.

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.

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