background Protocols

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Removal of the 5'-phosphate groups from the internal termini of bacteriophage {lambda} arms is used to prevent self-ligation and suppress the background of nonrecombinant bacteriophages during cloning. - [Read Alkaline Phosphatase Treatment of Bacteriophage lambda Vector DNA Protocol]

Category: Protein Protocols: Immunoprecipitation Protocols

Antibody-antigen complexes are removed from solution by addition of an insoluble form of an antibody binding protein such as Protein A, Protein G or second antibody. Immunoprecipitation protocols / methodology and technical background information. P.J. Ha - [Read Analysis of Proteins by Immunoprecipitation]

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocols for blocking solutions. Includes: Endogenous peroxidase blocking solution (H2O2-Azide); Non-specific antibody background blocking solution: swine serum; Non-specific antibody background blocking solution: MILK; Enzymatic antigen retrieval solution; Unconjugated, biotin- and fluorochrome-conjugated antibody (primary) solutions; Unconjugated, biotin- and fluorochrome-conjugated antibody (secondary) solutions; Enzyme-conjugated secondary antibody solutions.; etc.. - [Read Blocking Solutions Protocols]

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

Includes Abbreviations, Background, and Procedure steps using BSA. The Bradford protein assay (1) is one of several simple methods commonly used to determine the total protein concentration of a sample.  The method is based on the proportional binding of the dye Coomassie to proteins. The assay is colorimetric; as the protein concentration increases, the color of the test sample becomes darker.  Coomassie absorbs at 595 nm.  - [Read Bradford Protein Concentration Assay]

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]

Category: Microscopy Protocols: Optical Microscopy Protocols

The visibility of the faint star light is enormously enhanced against a dark background. This principle is applied in darkfield (also called darkground) microscopy, a simple and popular method for making unstained transparent specimens clearly visible. Such objects often have refractive indices very close in value to that of their surroundings and are difficult to image in conventional brightfield microscopy. - [Read Darkfield Illumination]

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol describes a denaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. The use of denaturing IPs reduces background considerably. - [Read Denaturing Protein Immunoprecipitation from Mammalian Cells Protocol]

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Immunoprecipitation Protocols

The use of denaturing Ips reduces background and simplifies protein harvest. Yeast are boiled in an SDS-containing solution to liberate total proteins. - [Read Denaturing Protein Immunoprecipitation from Yeast Protocol]

Category: Radioactivity

Radioactive Decay, Methods of detection, Liquid scintillation counting, Background and Quenching, The scintillation counter printout. David R. Caprette, Rice Univ. - [Read Detection and Measurement of Radioactivity]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Flow Cytometry Analysis Protocol. Springer Lab - Harvard. Background on flow cytometry technique: Flow cytometry is a method which uses instrumentation scanning single cells flowing past excitation sources in a liquid medium.
- [Read Flow Cytometry Analysis Protocol]

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

Information on Immortalized Mammary Epithelial Cells (IMEC). Includes: Background Information on IMEC; Culturing; Trypsinization. - [Read IMEC Maintenance]

Category: Molecular Imaging: Immunofluorescence Microscopy

Immunofluorescence staining. Includes Background, Procedure, Key points and materials / methods. Grainger Lab. - [Read Immunofluorescence staining]

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Immunoprecipitation Troubleshooting Guide. Stressgen. Problem: Non-specific background, Specific Background, Specific antigen not detected. With Solutions to common IP problems. - [Read Immunoprecipitation Troubleshooting Guide PDF]

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Immunoprecipitation TroubleShooting Guide. Sigma Aldrich. Problems: No binding, High background or unwanted proteins precipitate. With Solutions. - [Read Immunoprecipitation TroubleShooting Guide Sigma]

Category: Antibody Protocols: Antibody Labeling Protocols

Direct labeling of purified antibodies is the method of choice when simultaneously visualizing two or more antibodies of the same species, class, or subclass. This allows the localization of multiple antigens to be compared in the same cell, tissue, or sample. Labeled primary antibodies are also useful for improving background-to-readout ratios, and they can be essential for immunoassays in which good quantification is needed. - [Read Labeling Antibodies with Fluorochromes Protocol]

Category: Bacterial Protocols

Ice tea has a complex composition, which leads to reduced filterability, and a decrease in sample throughput. Its composition can generate background or false positive signals. It is also well known that ice tea contains molecules that can inhibit the bioluminescence reaction, which can generate false negative results. The aim of this study was to develop a protocol that was able to neutralize these affects and enable faster detection of contamination. - [Read Microbial Detection in Ice Tea Using the Millipore Milliflex Rapid Microbiology Detection System]

Category: Molecular Imaging: Immunofluorescence Microscopy

Minimizing Background in Immunofluorescence Procedures. Includes tips and methods on reducing background staining. Janis V. Giorgi Flow Cytometry Laboratory, UCLA. - [Read Minimizing Background in Immunofluorescence Procedures]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Immunofluorescence Staining Protocols

Minimizing background in immunofluorescence procedures. - [Read Minimizing Background in Immunofluorescence Procedures]

Category: Immunology

Minimizing background in immunofluorescence procedures. - [Read Minimizing Background in Immunofluorescence Procedures]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The MTT Cell Proliferation Assay measures the cell proliferation rate and conversely, when metabolic events lead to apoptosis or necrosis, the reduction in cell viability. The number of assay steps has been minimized as much as possible to expedite sample processing. The MTT Reagent yields low background absorbance values in the absence of cells. Includes: Determining optimal cell counts, performing an assay, data interpretations and troubleshooting. - [Read MTT Cell Proliferation Assay Protocol]

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

Using excitation at 365 nm and measuring emission at 455 nm, the amount of 4-MU produced can be quantified. Under these conditions, background fluorescence from the substrate is negligible, especially if the appropriate filter is selected. - [Read Quantitative GUS Activity Assay of Plant Extracts]

Category: Real Time PCR

Real-Time PCR Guide - Informative Background. M.Tevfik Dorak. - [Read Real-Time PCR Guide - Informative Background]

Category: PCR Protocols: Real-Time PCR Protocols

Protocol uses FAM-(6-carboxy-fluorescein) or JOE-(6-carboxy-4', 5' -dichloro-2',7' -dimethoxy-fluorescein) labeled LUX (Light Upon eXtension) primers, which can quantify 100 or fewer copies of the target DNA in a background of nonspecific templates, over a broad dynamic range of less than 100-107 copies.  It uses uracil deglycosylase (UDG) to minimize the risk of carryover contamination, and includes a melting curve analysis of the product. - [Read Real-Time PCR Protocol]

Category: Molecular Biology Protocols: Restriction Enzyme Digestion

Good background. Restriction Digestion of Lambda DNA. Dr. E.T. Wurtzel, BIO 420. - [Read Restriction Digestion for DNA]

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol for sizing of YAC clones. To determine the size of a yeast artificial chromosome within the background of the normal yeast chromosomal complement. - [Read Sizing of YAC Clones Protocol]