film Protocols

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

LCM isolates specific cells or tissues from samples mounted on microscope slides. The samples are viewed through a thermoplastic film that is attached to a microcentrifuge tube lid. Localized heat, caused by the application of a laser pulse, fuses the membrane to the cells of interest, which can then be harvested for further analysis. RNA and proteins can be purified from the isolated cells, allowing detailed analysis of gene expression. This protocol is divided into three stages. - [Read (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel]

Category: Radioactivity: Autoradiography

Autoradiography Protocols. Exposing gels and plates containing radioactive samples to X-ray film. Detection of 3H. Detection of 35S and 14C. Detection of 32P and 125I. Bart's Cookbook. - [Read Autoradiography Protocols]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Protocol for cloning genes from a phage library. Includes: Titer and plate out phage; Lift plaques onto filters and prepare them for screening; Make a probe; Hybridize the probe to the filters; Wash the filters and expose to film; Purify putative plaques; Excise plasmid from the desired phage. - [Read Clone Genes From a Phage Library Protocol]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to do film and digital imaging of microscopic preparations. - [Read How to Do Film and Digital Imaging of Microscopic Preparations]

Category: Antibody Protocols: Antibody Labeling Protocols

This protocol describes a simple chemical oxidation method for labeling antibodies with iodine. Iodide-125 (supplied as NaI) is oxidized to form iodine-125 (I2), which attacks tyrosyl and histidyl side chains. The iodinated antibodies are easily detected and quantitated using gamma counters or film. They are used primarily in immunoassays, but other techniques can be adapted conveniently to the iodine detection method. - [Read Labeling Antibodies with Iodine Protocol]

Category: Gene Delivery Protocols: DNA Nanoparticles Protocols

Lipoplex (cationic liposome-DNA complex) is formed via electrostatic interaction of anionic nucleic acids with cationic liposomes. A thin film of lipids is dried on the bottom of a glass tube and rehydrated in an aqueous solution. The resulting liposome suspension is passed through polycarbonate filters of desired pore size. This protocol also describes the preparation, physical properties, and biological activity of liposome-polycation-DNA (LPD) nanoparticles. - [Read Lipoplex and LPD Nanoparticles for In Vivo Gene Delivery Protocol]

Category: Protein Protocols: Western Blot Protocol: Troubleshooting Western Blots

Troubleshooting Guide for Chemiluminescent Western Blotting Pierce. High Background, Reverse image on film, Blot glows in the darkroom, Signal duration is less than 8 hours, Weak or no signal, Spots within the protein bands, non-specific bands. - [Read Troubleshooting Guide for Chemiluminescent Western Blotting Pierce]

Category: Protein Protocols: Western Blot Protocol: Troubleshooting Western Blots

Streaking of Western Blots, Weak Bands or Weak Staining of Western Blots, Poor Transfer of Membranes, Dirty Blot, Multiple Bands on Western Blot Membrane and Film. Western Blot Info. - [Read Western Blot Troubleshooting]