Protein Microarray Chips - An Introduction. Introduction, Types of Protein Chips, Attachment, Protein and Antibody Chip Production, Applications of Protein Chips, Detection methods, and Future Directions.
Molecularstation. - [Read Protein and Antibody Microarray Chips - An Introduction]
Western Blot Home- http://www.molecularstation.com/protein/western-blot/
Western Blot Home. Information on western blotting, western blot procedure and methods, western blot books, stripping protocols. - [Read Western Blot Home]
Covers Many Western Blot Problems and Includes Many Solutions. Fuzzy Bands, Low or Weak Signal, High Background, Spots on Film, Too Many Bands. A MolecularStation Guide. - [Read Troubleshooting Western Blot]
Immunostaining thin layer chromatograms TLC is a very sensitive detection technique of functionally active carbohydrate ligands of protein receptors. Carbohydrate structures are detected in glycolipids from complex mixtures of molecules extracted from the relevant target tissue. Proteins analyzed can be antibodies, chimeric Ig proteins, selectins, lectins, toxins, and other carbohydrate binding proteins. John L. Magnani~GlycoTech Corporation, Rockville, Maryland - [Read Immunostaining Thin Layer Chromatograms Of Glycolipids]
In Vitro Footprinting Methods Protocol- http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/5.%20In-vitro%20footprinting.pdf
Protocol for in vitro transcription and translation using the coupled reticulocyte lysate system. This protocol is designed to test random samples on a protein gel. Scale up the reactions accordingly. Protocol includes: Procedure, Solutions, BioReagents and Chemicals and protocol hints. - [Read In Vitro Transcription and Translation Using the Coupled Reticulocyte Lysate System]
In-Gel Digestion Protocol. Excision of protein bands (spots) from polyacrylamide gels Reduction and alkylation . Reduction and alkylation . MALDI analysis of the supernatant after in-gel digestion. Extraction of Peptides.Matthias Wilm EMBL Bioanalytical R - [Read In-gel digestion of proteins to be analyzed by mass spectrometry]
Activation and inactivation of proteins using photoactivation of caged peptides or proteins offer insights into cellular dynamics not achievable using genetic means. The ability to selectively alter the activity of a specific protein at a defined time and location inside a cell allows the correlation of changes in protein activity and cellular behavior. A caged compound, peptide, or protein is prepared by covalently linking it to a photolabile, protecting group. - [Read Introduction of Caged Peptide/Protein into Cells Using Microinjection Protocol]
Live-cell imaging techniques provide critical insight into the fundamental nature of cellular & tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein & synthetic fluorophore technology. Because of these advances, live-cell imaging has become a requisite analytical tool in most cell biology labs. Includes: Maintaining Live Cells on the Microscope Stage; Live-Cell Imaging Culture Chambers; Optical System and Detector Requirements etc. - [Read Introduction to Live-Cell Imaging Techniques]
The basis of this procedure is that two specific cell type preparations may be isolated, exposed separately to various compounds over a range of concentrations, and the cytotoxicity of these determined. Parameters deemed indicative of a cytotoxic effect include a reduction in de novo protein synthesis and decreased glucose and fatty acid metabolism. A cytotoxic effect may indicate that a chemical is likely to be nephrotoxic in vivo. - [Read Isolated Rat Glomeruli and Proximal Tubules]
Isolation of proteins from inclusion bodies Protcol. Expression of recombinant proteins. Dissolving of recombinant protein. Refolding buffer. BJÖRKman group - [Read Isolation of proteins from inclusion bodies]
Protocol describes how tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay. Protocol includes: Translation of Xenopus Mos Kinase; Antibody to Antigen Binding; Protein A Sepharose to Antibody Binding; Kinase Reactions on Immunoprecipitated Material; Polyacrylamide Gel Analysis of Immunoprecipitates. Includes protocol hints. - [Read Kinase Assay Using In Vitro Translated Xenopus Mos Kinase]
Two methods are provided for purifying glycoproteins using wheat-germ agglutinin or concanavalin A-Sepharose. Because lectin-affinity matrices can bind a few milligrams of protein per milliliter of affinity matrix, only a small amount of affinity gel matrix is required. The batchwise method is recommended when protein volume is large. - [Read Lectin-Agarose Affinity Chromatography Protocol]
Find a list of assays for the determination of protein concentration in a solution. This list includes the sensitivity range, volume/amount of sample needed, subjective comments on accuracy and convenience, and major interfering agents. Procedural details, equipment requirements, and references are outlined in the individual assay documents. - [Read List of Protein Assays]
Protocol describes the acquisition and processing of confocal
fluorescent and bright field images of live cells, expressing cyan fluorescent protein(CFP) and/or yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M microscope. This procedure is used to help determine if N- or Cterminal tagging of signaling molecules alters the steady state localization pattern of the signaling protein in question. - [Read Live Cell Spinning Disk Confocal Fluore Imaging of Cells- Colocalization of Fluorescent Protein Tags]
Protocol describes the acquisition and processing of confocal
fluorescent and bright field images of live cells expressing yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M microscope when three planes along the z-axis of the cell are acquired. Protocol includes: Description of Microscope and Imaging Setup; Description of Acquisition Parameters; Image Processing. - [Read Live Cell Spinning Disk Confocal Fluorescence Imaging of Cells- YFP & Bright Field—Three Z Axis]
Protocol describes the acquisition and processing of confocal
fluorescent and bright field images of live cells expressing yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M microscope. Protocol includes: Description of Microscope and Imaging Setup; Description of Acquisition Parameters; Image Processing. - [Read Live Cell Spinning Disk Confocal Fluorescence Imaging of Cells- YFP and Bright Field Images]
Protocol describes the acquisition and processing of confocal
fluorescent images of live cells expressing yellow fluorescent protein (YFP), with a spinning disk confocal head on a Zeiss Axiovert 200 M microscope. Protocol includes: Description of Microscope and Imaging Setup; Description of Acquisition Parameters; Movie Processing. - [Read Live Cell Spinning Disk Confocal Fluorescence Imaging of Cells- YFP Time Series for Markers]
Lowry – Protein Determination. Quantitation of total protein using the lowry protein assay. Dr. Mario Lebendiker. The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem. Method materials and notes for the Lowry assay. - [Read Lowry – Protein Determination]
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]
Lowry Protein Assay Protocol. Yu-li Wang Lab, Univ. Massachusetts Medical School. Detailed protocol including materials. - [Read Lowry Protein Assay Protocol]
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)]
Potential embryotoxicity is assessed by monitoring the effect of the test compound on total protein synthesis, and DNA synthesis in cultured human foetal lung fibroblasts. Rat lung epithelial cells can be used to determine cytotoxicity of select compounds because of their ability to metabolise xenobiotics. - [Read Lung Cell Assay Protocol]
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 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 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.
A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.
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.