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Western Blot Home - http://www.molecularstation.com/protein/western-blot/

Category: Protein Protocols: Western Blot Protocol

Western Blot Home. Information on western blotting, western blot procedure and methods, western blot books, stripping protocols. - [Read Western Blot Home]

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Low-melt Polyester Embedding Protocol - http://cgap-mf.nih.gov/Protocols/Tissues/TissueProtocols/LowMeltPolyester.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols: Tissue Preparation for LCM Protocols

Protocol for low-melt polyester embedding. Includes: Large Tissue Specimens; Small Tissue Specimens and Biopsies; Histology Slide Cutting Procedure. - [Read Low-melt Polyester Embedding Protocol]

Lowry Protein Assay - http://www.animal.ufl.edu/hansen/protocols/LOWRY.htm

Category: Protein Protocols: Lowry Protein Assay

Lowry Protein Assay. The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-275 (1951)]. Under alkaline conditions, copper complexes with protein. When folin phenol reagent (phospho-molybdic-phosphotungstic reagent) is added, the Folin-phenol reagent binds to the protein. Bound reagent is slowly reduced and changes color from yellow to blue. P.J. Hansen, Dept. of Animal Sciences, University of Florida. - [Read Lowry Protein Assay]

Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000162.pdf?pid=PP00000162

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol is the first step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LC-MS/MS. This procedure is used to prepare protein extracts from WEHI-231 cells. This preparation method provides total cellular protein samples that are free of contaminating nucleic acids. - [Read Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol]

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)]

Markers for Pulsed-field Gel Electrophoresis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3222

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol fopr markers of pulsed-field gel electrophoresis. Markers for pulsed-field gel electrophorsis can be generated by ligation of linear monomers of bacteriophage {lambda} DNA (48.5 kb) into a nested series of concatemers. This procedure yields a series of concatemers that contain up to 20 tandemly arranged copies of bacteriophage DNA. - [Read Markers for Pulsed-field Gel Electrophoresis Protocol]

MEF Mouse Embryonic Fibroblast Protocols - http://ink.primate.wisc.edu/~thomson/protocol.html#harvest

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

MEFs. Thompson Lab Stem Cell Methods and Procedure. Irradiation and plating MEFs, Splitting MEF cells, Thawing MEF, Freezing and Harvestion MEF cells. - [Read MEF Mouse Embryonic Fibroblast Protocols]

Methanol-Chloroform Precipitation of Proteins - http://www.healthsystem.virginia.edu/internet/biomolec/methanolchloroform.cfm

Category: Mass Spectrometry Protocols

Methanol-Chloroform Precipitation of Proteins. A procedure for precipitating proteins from solution, including detergent solutions, is from Wessel and FlÃ¼gge (1984). Univ. Virginia. - [Read Methanol-Chloroform Precipitation of Proteins]

Method: Preparation of Lymphocyte Cell Pellet for Storage - http://humgen.wustl.edu/hdk_lab_manual/hcc/hcc5.html

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

Following propagation to 1 X 108 cells, lymphoblastoid cells are conveniently stored at -80 degrees C to preserve the high molecular weight DNA in the cells until the DNA is purified. This procedure describes the steps required to harvest and freeze the c - [Read Method: Preparation of Lymphocyte Cell Pellet for Storage]

Methods for Maintaining Insect Cell Cultures - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=355909

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Insect cell cultures are now commonly used in insect physiology, developmental biology, pathology, and molecular biology. As the field has advanced from methods development to a standard procedure, so has the diversity of scientists using the technique. This paper describes methods that are effective for maintaining various insect cell lines. The procedures are differentiated between loosely or non-attached cell strains, attached cell strains, and strongly adherent cell strains. - [Read Methods for Maintaining Insect Cell Cultures]

Microarray Protocol for Affymetrix In Situ Synthesized Oligo Arrays - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000174.pdf?pid=PP00000174

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol details the preparation of biotin-labeled target samples and hybridization of these samples to an Affymetrix in situ synthesized oligonucleotide GeneChip array. The procedure requires a minimum of 5 µg of purified total RNA as starting material. - [Read Microarray Protocol for Affymetrix In Situ Synthesized Oligo Arrays]

Microarray Protocol for Agilent Inkjet-Deposited Presynthesized - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000184.pdf?pid=PP00000184

Category: Microarray Protocols: Amino-allyl Microarray Methods

Protocol details the preparation of fluorescently labeled target samples (aminoallyl method) and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides. The procedure requires a minimum of 3 µg of purified total RNA as starting material. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized]

Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Array - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000179.pdf?pid=PP00000179

Category: Microarray Protocols: Microarray DNA Labeling Protocols

Protocol details the preparation of fluorescently labeled target samples and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides. The procedure requires a minimum of 3 µg of purified total RNA as starting material. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Array]

Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Arrays - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000179.pdf?pid=PP00000179

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol details the preparation of fluorescently labeled target samples and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides. The procedure requires a minimum of 3 µg of purified total RNA as starting material. Includes: cDNA Synthesis; Fluorescent cRNA Synthesis; cRNA Precipitation and Cleanup; cRNA Quantification; Hybridization; Washing. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Arrays]

Monoclonal Antibody Production Fusion Protocol - http://www.icp.ucl.ac.be/mexp/vipg_fus.html

Category: Antibody Protocols: Antibody Production Protocols

Protocol for monoclonal antibody production fusion. Fusion is an important procedure to produce monoclonal antibodies (MoAb). - [Read Monoclonal Antibody Production Fusion Protocol]

Nitrocellulose Membrane Washing Procedure for cDNA Screening - http://www.biochem.uiowa.edu/donelson/Database%20items/cDNA_Scrn_Washing.htm

Category: RNA Protocols: cDNA Libraries Library

Denaturation Solution, Neutralization Solution, Rinse Solution, Crosslink DNA to membranes using Stratalinker UV crosslinker. Donelson Lab, Univ. Iowa. - [Read Nitrocellulose Membrane Washing Procedure for cDNA Screening]

Opening the Abdominal Cavity and Locating Female Reproductive Organs Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4356

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes the cervical dislocation method for humane sacrifice of a mouse, as well as a technique for dissecting the female reproductive organs. Local institutional regulations should be followed for this procedure. - [Read Opening the Abdominal Cavity and Locating Female Reproductive Organs Protocol]

Optimized Protocol for Mounting Tissue Sections PDF - http://arctur.com/research_portal/images/Application%20Notes/pen_membrane_protocol_9.pdf

Category: Histology Protocols: Tissue Sample Sectioning

Generally Required Materials and Equipment for Tissue Sectioning. Sectioning and Slide Preparation Procedure onto Metal-Framed PEN Membrane Slides. Arcturus. - [Read Optimized Protocol for Mounting Tissue Sections PDF]

Ovariectomy for Induction of Blastocyst Implantation Delay Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4378

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes a method for removal of mouse ovaries in combination with the administration of progesterone, which causes the arrest of blastocyst development. This procedure is performed during the afternoon of the third day of pregnancy to ensure that the morulae have moved sufficiently into the oviducts and will be less likely to be damaged during the ovariectomy. - [Read Ovariectomy for Induction of Blastocyst Implantation Delay Protocol]

Oviduct Transfer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4379

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes oviduct transfer of mouse embryos. It is based on the Whittingham method, which applied a well described procedure for the rat to the mouse. It is best to practice this procedure first on a cadaver and then on an anesthetized 0.5-dpc pseudopregnant mouse using a dye solution or blue Affigel beads rather than embryos to gain experience in finding the opening of the oviduct (infundibulum). - [Read Oviduct Transfer Protocol]

Packing an IMAC Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4215

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol for packing an IMAC column. Protocol describes a procedure for packing a 30-ml immobilized metal-ion affinity chromatography (IMAC) column with the aid of a pump. - [Read Packing an IMAC Column Protocol]

Packing Capillary Columns for RP-HPLC Protocol - http://cshprotocols.cshlp.org/cgi/content/abstract/2006/28/pdb.prot4546

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 µl/min) and gradients required to operate slurry-packed capillary columns. A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to a number of commercial UV detectors. - [Read Packing Capillary Columns for RP-HPLC Protocol]

PAR-1 Staining Procedure Protocol - http://www-biology.ucsd.edu/labs/aroian/protocols/PAR1-stain.htm

Category: C. elegans Protocols: C. elegans Staining Protocols

Protocol for PAR-1 staining procedure. Includes fixation and staining. - [Read PAR-1 Staining Procedure Protocol]

Paraffin Sectioning for LCM Plant Tissue - http://schnablelab.plantgenomics.iastate.edu/docs/resources/protocols/Paraffin_sectioning_for_LCM.pdf

Category: Histology Protocols: Tissue Sample Sectioning

FIXATION, DEHYDRATION, PARAPLAST INFILTRATION AND SECTIONING PROTOCOL WITH TISSUE COLLECTION PROCEDURE USING THE PALM LASER MICRODISSECTION SYSTEM. Schnable Labs, Plant Genomics. - [Read Paraffin Sectioning for LCM Plant Tissue]

Passage Procedure for RAW 264.7 Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000159.pdf?pid=PP00000159

Category: Cell Culture Protocols: Cell Lines Protocols

RAW 264.7 cells are a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice. For routine maintenance in culture (passage), cells are seeded at a confluence of approximately 10% (1 x 106 and 3 x 106 cells in 100-mm and 150-mm plates, respectively) and grown to a confluence of approximately 80%. This procedure requires the cells to be split every two days. - [Read Passage Procedure for RAW 264.7 Cells]

PCR Amplification of Highly GC-Rich Regions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4093

Category: PCR Protocols: PCR Optimization

Procedure details the establishment of an amplification procedure for GC-rich sequences. The DNA fragments of interest are amplified in the presence of either 5% DMSO, 1 M betaine, 2 M betaine, 1 M betaine, and 5% DMSO; 2 M betaine and 5% DMSO; 0.4 M tetramethylene sulfone; or without any of the enhancers. - [Read PCR Amplification of Highly GC-Rich Regions Protocol]

Articles (1-10 of 12)

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.

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.