native Protocols

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

Protocols on band shift. Includes: Phosphorylation of recombinant antibodies; Labeling the antigen with a red fluorophore; Native gel electrophoresis; Band-shift assay for the determination of Kd; Band-shift assay for the determination of koff. - [Read Band Shift Protocols]

Category: Plant Biology Protocols: Plant Proteins, Peptides and Antigens

BN-PAGE has become the method of choice for the investigation of the respiratory protein complexes of the electron transfer chains of a range of organisms. It allows the separation in two dimensions of extremely hydrophobic protein sets for analysis and also provides information on their native interactions. In this review we discuss the capabilities of BN-PAGE in proteomics and the wider investigation of protein:protein interactions with a focus on its use and potential in plant science. - [Read Blue-Native PAGE in Plants: A Tool in Analysis of Protein-Protein Interactions]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocols for gene expression and protein localization in Arabidopsis. Includes: Detection of the native protein; Detection of a recombinant version; Immunofluorescence detection in Arabidopsis protoplasts; Isolation of Arabidopsis seedling protoplasts; Subcellular localization of GUS-fusion proteins in Arabidopsis seedlings; Localization of Arabidopsis proteins with GUS in situ enzyme assay. - [Read Gene Expression and Protein Localization in Arabidopsis Protocols]

Category: Microscopy Protocols

The atomic force microscope (AFM) is one of the most powerful tools for determining the surface topography of native biomolecules at subnanometer resolution. The AFM can also provide insight into the binding properties of biological systems. In order to determine the specific interaction between two kinds of molecules (e.g., avidin and biotin). Includes information on principle of AFM and application of AFM. - [Read Imaging, Measuring and Manipulating Native Biomolecular Systems with the Atomic Force Microscope]

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with anti-phosphotyrosine:biotin conjugates. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation with Anti-Phosphotyrosine:Biotin Conjugates Protocol]

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with antibody:agarose conjugates. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation with Antibody:Agarose Conjugates Protocol]

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation with soluble antibodies. Includes: Denaturing Conditions; Native Conditions; Pre-clearing. - [Read Immunoprecipitation With Soluble Antibodies Protocol]

Category: Microscopy Protocols: Live-Cell Imaging Protocols

GFP serves as a molecular marker that can be imaged dynamically in living cells, both in its native form & as a fusion to other proteins. For GFP imaging, plants present the challenge of autofluorescence from chlorophyll, lignified cell walls, vacuolar contents, and other cell materials, all of which can obscure the GFP signal. Maximizing the signal-to-noise ratio is a major concern, and careful consideration should be given to the choice of tissue imaged, GFP expression level, etc. - [Read Live-Cell Imaging of GFP in Plants]

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

Native Aequorin. NanoLight Technology. Aequorin has advantages over other Ca2+ indicators, for example, low leakage rate from cells, lack of intracellular compartmentalization or sequestration and it does not disrupt cell functions or embryo development. - [Read Native Aequorin]

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Native Chromatin Immunoprecipitation Protocol ABCAM - [Read Native Chromatin Immunoprecipitation Protocol]

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes a nondenaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. However, the nondenaturing protocol is useful when one wishes to separate soluble from insoluble proteins, or when the antibody being used recognizes a native epitope. - [Read Nondenaturing Protein Immunoprecipitation from Mammalian Cells Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

An in vitro red blood cell assay is presented which allows the estimation of the irritation potential of tensides and tenside containing materials such as shampoos, shower gels, cleaning products, etc. The estimation is based on the fact that surfactants interact strongly with cellular membranes and proteins.  Both effects are measured photometrically by use of the inherent native dye, oxyhemoglobin. - [Read Red Blood Cell Test System Protocol]

Category: Microscopy Protocols: Optical Microscopy Protocols

The light microscope allows dynamic biological processes to be imaged in their native (i.e., aqueous) environment with relatively high temporal resolution. However, the diffraction-limited resolution is low. When working at or beyond the diffraction-limited resolution of the LM, a disadvantage of fluorescence imaging is the relatively low signal-to-noise (S/N) ratio of the images. However, this can be increased significantly by video and computer technology. - [Read Watching Molecular Motors at Work by Video-Enhanced Light Microscopy]