Monday, May 10, 2004 Description WHOLE BLOOD LYMPHOCYTE PROLIFERATION ASSAY Procedure PRINCIPAL AND/OR PURPOSE OF THE TEST Lymphocyte proliferation assays (LPA) have been widely used to help describe the abnormalities associated with diver...

Monday, May 10, 2004 Description RESPONDER CELL FREQUENCY ASSAY (RCF): LPA Limiting Dilution Procedure 1 This assay measures the frequency of pathogen-specific memory CD4 cells (1) by adding a limiting dilution step to a lymphocyte prolife...

Monday, May 10, 2004 Description Yeast IF without Dehydration Procedure Grow cells at the appropriate temperature to 5x10E6 in 5 mls YPD. Add 0.5mls 37% formaldehyde (best grade) and incubate on the roller at same temp. for 10 min. Spin do...

Monday, May 10, 2004 Description Yeast IF with Methanol/Acetone Dehydration Procedure Grow cells at the appropriate temperature to 5x10E6 in 5 mls YPD. Add 0.5mls 37% formaldehyde (best grade) and incubate on the roller at same temp. for 1...

Monday, May 10, 2004 Description This protocol will visualize both internalized receptors that were once on the plasma membrane and receptors in the biosynthetic pathway. This protocol was specifically written for visualizing agonist-induc...

Monday, May 10, 2004 Description This protocols allows the use of fluorescence microscopy to visualize epitope-tagged receptors expressed by stable adherent cell lines. In contrast to the permeabilized cell protocol, this protocol where an...

Monday, May 10, 2004 Description The Following Protocol can be used to couple either FITC or Biotin to IgG. The same method is used for both, and when discrepancies occur, they will be noted. Procedure . Dissolve 2mg of IgG in 1mL of PBS (...

Monday, May 10, 2004 Description PDS1::HA IMMUNOFLUORESCENCE Procedure Cultures: 1500, 1501 Fixations: 10, 20, 30 and 45 minutes in 4% formaldehyde, 0.1 M KPO4 pH 6.5; 15 minutes in 4% formaldehyde, 50 mM KPO4 pH 6.5, 0.5 mM MgCl2 Grow cel...

Monday, May 10, 2004 Description IIF Protocol (modifications for maintaining GFP fluorescence Procedure Grow cells to OD 0.5 Harvest 0.5-1 ml cells. Resuspend in PBS, and add PFA to final concentration of 4%. If using the regular formaldeh...

Monday, May 10, 2004 Description Iindirect immunoflourescence for yeast: Procedure 1mL yeast (~0.2-0.5 OD) + 0.1mL 36% formaldehyde, 60'/Rt Wash 2x 1mL H2O (spin 10K, 5). Wash in 1mL SK and resuspend in 450 ul SK. Sonicate 5 sec. lowest se...