Thursday, October 16, 2003 Description This method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to enter the cells. Propidium Iodide (PI) is a DNA-binding fluoroch...
Thursday, October 16, 2003 Description This is the method for direct immunofluorescence labeling; that is, the antibodies have the fluorescent dye attached. Direct labeling is simpler and quicker than indirect labeling. Procedure Adjust th...
Thursday, October 16, 2003 Description Procedures for cell plating, fixing, antibody incubation and mounting Procedure A. Plating: To sterilize glass coverslips, dip in ethanol and flame. We use 22x22x1 mm3 coverslips and put them in 6-wel...
Tuesday, March 01, 2005 Description This is the standard Ellmans Test for the determination of free thiols. (Ref. 1) It works well for small peptides and proteins synthesized using standard solid phase synthetic methods. Peptides from thes...
Monday, June 07, 2004 Description This rubidium chloride protocol gives better transformation efficiencies than calcium chloride procedures for most strains. Procedure 1) Innoculate a single colony from an LB plate in 2.5 ml of LB medium....
Wednesday, June 02, 2004 Description This protocol explains how to prepare Magnesium/Calcium competent cells for transformation with DNA. It is described using BL21 cells but other E. Coli cells could be used. Procedure 1: Innoculate a sin...
Wednesday, June 02, 2004 Description This method can be use to quickly identify if the correct plasmid (identified here by size) has been transformed into a bacterial cell (generally E. Coli). Procedure 1: Grow bacterial colonies on rich a...
Wednesday, May 05, 2004 Description Post-Embedding Immunogold Method Procedure L.R. white sections are collected on gold Veco grids. If blocking is necessary for nonspecific labeling, buffer is supplemented with normal goat serum (2 - 10%)...
Wednesday, May 05, 2004 Description Immunofluroescence Technique Procedure Fix cells in 2% formaldehyde in PBS/pH 7.4 for 15 min. at 20oC. 2% formaldehyde is made up fresh prior to use by dissolving the appropriate amount of EM grade paraf...
Monday, May 03, 2004 Description Preparing Serum from Whole Blood Procedure Put the blood at 37 for 30-60 min to clot. 3. Pull the rubber stopper off the top, and use a long Pasteur pipette to ring the clot, i.e. separate it from the sides...