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A Method for Assaying Deubiquitinating Enzymes - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=140113

Category: Protein Protocols: Protein Ubiquitination Protocols

A Method for Assaying Deubiquitinating Enzymes. Lee et al., Biol Proced Online. May 1998. A general method for the assay of deubiquitinating enzymes was described in detail using 125I-labeled ubiquitin-fused Î±NH-MHISPPEPESEEEEEHYC (referred to as Ub-PESTc) as a substrate. - [Read A Method for Assaying Deubiquitinating Enzymes]

Amplification of cDNA Generated by Reverse Transcription of mRNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3837

Category: PCR Protocols: cDNA Synthesis Protocols

An oligodeoxynucleotide primer hybridized to mRNA is extended by an RNA-dependent DNA polymerase to create a cDNA copy that can be amplified by PCR. Depending on the purpose of the experiment, the primer for first-strand cDNA synthesis can be specifically designed to hybridize to a particular target gene, or a general primer such as oligo(dT) can be used to prime cDNA synthesis from essentially all mammalian mRNAs - [Read Amplification of cDNA Generated by Reverse Transcription of mRNA Protocol]

Annexin V Staining Flow Cytometry Protocol - http://www.cyto.purdue.edu/flowcyt/research/cytotech/amfc/data/page1.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Annexin V Staining Flow Cytometry Protocol - a general protocol for staining cell for cytometry analysis. BD PharMingen. - [Read Annexin V Staining Flow Cytometry Protocol]

Bradford Assay Method - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Biochemistry/Bradford.html

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

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose.
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.

Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]

Constructing and Expressing GFP Fusion Proteins - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4649

Category: Reporter Gene Assays: GFP Assay Protocols

The following protocol can be used for the development of stable cell lines expressing GFP fusion proteins. Although optimal transfection procedures (e.g., calcium phosphate, electroporation, or FuGENE 6 [Roche Applied Science]) vary depending on cell type, this general transfection procedure has been successful for stable transfection of HeLa, A-431, U2OS, BHK, and HT1080 cells. - [Read Constructing and Expressing GFP Fusion Proteins]

Contrast in Optical Microscopy - http://microscopy.fsu.edu/primer/techniques/contrast.html

Category: Microscopy Protocols: Optical Microscopy Protocols

When imaging specimens in the optical microscope, differences in intensity and/or color create image contrast, which allows individual features and details of the specimen to become visible. Contrast is defined as the difference in light intensity between the image and the adjacent background relative to the overall background intensity. In general, a minimum contrast value of 0.02 (2 percent) is needed by the human eye to distinguish differences between the image and its background. - [Read Contrast in Optical Microscopy]

Cytochrome P450 Recombinant Enzyme Microsome Protocol - http://www.researchd.com/cyp450/microsom.htm

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol for Cytochrome P450. Includes: Microsome Assays General; General Reaction Conditions; Specific Substrates for Cytochrome P450 Isozymes. - [Read Cytochrome P450 Recombinant Enzyme Microsome Protocol]

ENLIGHTNING - General Procedure for Gel Autoradiography Enhancement - http://las.perkinelmer.com/applicationssummary/applications/enlightning+protocol.htm

Category: Radioactivity: Autoradiography

Gel Autoradiography Protocols. Drying the gel, exposure, and autoradiography. Protein Fixation and Staining (Optional). ENLIGHTNING - General Procedure for Gel Autoradiography Enhancement. PerkinElmer - [Read ENLIGHTNING - General Procedure for Gel Autoradiography Enhancement]

Epitope Tagging of Recombinant Proteins Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E663998FCCB61070F32F1EAE0301F7F&objectid=6677C749FA75FD435618AC07256968A7

Category: Immunology: Immunoprecipitation Protocols

Protocols for detection and purification of proteins tagged with a particular epitope, the FLAG tag, although the same general approach can be applied to other epitope tags. The protocols employ the anti-FLAG M2 antibody to detect and purify FLAG-tagged proteins. The methods presented are immunoprecipitation of FLAG fusion proteins from cells using an anti-FLAG M2 affinity gel, detection of FLAG fusion proteins by western blotting, and purification of FLAG fusion proteins by anti-FLAG. - [Read Epitope Tagging of Recombinant Proteins Protocol]

General Cell Culture - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/GeneralCulture.html

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Covers general things you need to know to get started in cell culture work. Includes: Sterile Technique; Growth of Your Cell Culture; - [Read General Cell Culture]

General Confocal Protocols - http://www.rci.rutgers.edu/~rsharma1/protocols.html

Category: Molecular Imaging: Confocal Microscopy Protocols and Information

General Confocal Protocols. Confocal Startup Protocol, Confocal Clean-Up Protocol, Confocal Printing Protocol.CONVENTIONAL MICROSCOPY NOTES. Piscataway, New Jersey. Rutgers University - [Read General Confocal Protocols]

General EMSA Protocol - http://faculty.virginia.edu/mirmira/Protocols/emsa.html

Category: DNA Protocols: EMSA DNA-Protein Protocols

There are multiple variations to this protocol, but they find that this one works well in all cases tested. Mirmira Lab. - [Read General EMSA Protocol]

General Guide for Cryogenically Storing Animal Cell Cultures - http://www.corning.com/lifesciences/technical_information/techdocs/cryoanimalcc.asp

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Advantages of Freezing Cell Cultures, Practical Aspects of Cell Freezing, Cryoprotection, Storage Vessels, Labeling and Recordkeeping, Recovery, Managing a Cell Repository, cell selection, Problem Solving Suggestions. Corning. by John A. Ryan - [Read General Guide for Cryogenically Storing Animal Cell Cultures]

General Immunofluorescence Protocol - http://www.bcm.edu/rosenlab/protocols/generalIF.pdf

Category: Molecular Imaging: Immunofluorescence Microscopy

General Immunofluorescence Protocol. Rosen Lab. Deparaffinize and rehydrate sections, Antigen retrieval methods, Proteinase K Antigen Retrieval, Urea Antigen Retrieval, Blocking buffers, - [Read General Immunofluorescence Protocol]

General Immunohistochemistry Protocol - http://www.bcm.edu/rosenlab/protocols/generalIHC.pdf

Category: Immunohistochemistry Protocols

Protocol for general immunohistochemistry. Includes: Antigen retrieval methods; Sodium Citrate Antigen Retrieval; Proteinase K Antigen Retrieval; Urea Antigen Retrieval. - [Read General Immunohistochemistry Protocol]

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]

General Overview of Histology - http://www-bioc.rice.edu/bios576/histology/histology.htm

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Fixation must convert the soluble contents of the cell (ie proteins, carbohydrates, ... http://www.protocol-online.org/prot/Histology/Fixation/ ...General Overview of Histology and Tissue Fixation. BIOS 576. Rice. - [Read General Overview of Histology]

General PCR Protocol - http://www.biology.lsa.umich.edu/research/labs/maddock/protocols/PCR/general_pcr_protocol.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

General PCR Protocol. Sally A. Green - [Read General PCR Protocol]

General PCR Protocol - http://www.biology.lsa.umich.edu/research/labs/maddock/protocols/PCR/general_pcr_protocol.html

Category: PCR Protocols

General PCR Protocol. Sally A. Green - [Read General PCR Protocol]

General PCR Protocol - Sally Green - http://www.biology.lsa.umich.edu/research/labs/maddock/protocols/PCR/general_pcr_protocol.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: Standard PCR Protocol

General PCR Protocol - Sally Green 1996 - [Read General PCR Protocol - Sally Green]

General PCR Protocol - Sally Green - http://www.biology.lsa.umich.edu/research/labs/maddock/protocols/PCR/general_pcr_protocol.html

Category: PCR Protocols: Standard PCR Protocol

General PCR Protocol - Sally Green 1996 - [Read General PCR Protocol - Sally Green]

General Principles of Immunoprecipitation - Great Tips - http://web.archive.org/web/20041125171142/http://flosun.salk.edu/~sefton/Hyper_protocols/immunoprecip.html

Category: Protein Protocols: Immunoprecipitation Protocols

Great Tips on IP - Bart's Cookbook - [Read General Principles of Immunoprecipitation - Great Tips]

General Stain for Toludine Blue Protocol - http://www.nottingham.ac.uk/pathology/protocols/tolblue.html

Category: Neuroscience Protocols: Histology Stains Protocols: Mucin Stains Protocols

Protocol for toludine blue general stain. - [Read General Stain for Toludine Blue Protocol]

GENERAL USE OF ENZYMES IN MOLECULAR BIOLOGY - http://www.people.hofstra.edu/faculty/Beverly_Clendening/Adv_Molecular_Biology/Protocols/General_use_of_Enzymes.html

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

GENERAL USE OF ENZYMES IN MOLECULAR BIOLOGY. Basics of Restriction Enzymes. - [Read GENERAL USE OF ENZYMES IN MOLECULAR BIOLOGY]

Hints and Precautions for the Care, Feeding and Breeding of Neurospora - http://www.fgsc.net/methods/perkins.html

Category: Fungi Protocols: Neurospora Protocols

Hints and precautions for the care, feeding and breeding of Neurospora. Includes: Crossing; Tips on handling ascospoes General Laboratory Practices; Stockkeeping; Mutagenesis and enrichment; Tips for encouraging colonial growth; Solutions and media; Handling Heterokaryons. - [Read Hints and Precautions for the Care, Feeding and Breeding of Neurospora]

Articles (1-9 of 9)

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.

Chrysoidin Stain

Chrysoidin Stain preparation method and protocol.

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.

Thionin Staining

Thionin Staining method.

Thionin Method

Thionin Method and protocol for staining cells.

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.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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