Friday, April 29, 2005 Description This assay allows the detection of cytolytic activity specific for a peptide epitope in vivo in mice. One group of cells is labelled with a fluorescent dye and pulsed with a peptide epitope. The other gro...
Wednesday, May 05, 2004 Description Protocol for intracytoplasmic staining of cytokines for FACS analysis Procedure 1) Prepare spleen, lymph node or T cell clone cells as single cell suspension, using erythrocyte lysis (water) or Lymphopaq...
Wednesday, May 05, 2004 Description This method is useful for monitoring depletion in vivo using rat IgG2b monoclonal antibodies against CD4 and CD8. The indirect staining method, using a monoclonal reagent that does not give any backgroun...
Monday, May 03, 2004 Description One way to eliminate crossreactivity of a polyclonal antiserum raised against a worm protein is to preabsorb the antiserum against total protein extracted from a worm mutant that does not produce that speci...
Monday, May 03, 2004 Description Protocol for Metal Enhanced DAB Staining using HRP 2 Antibodies Procedure 1. Carry out the fixation, permeabilization, and primary antibody incubation steps exactly as described above for the FITC detection...
Description This protocol works for many antibodies. A much longer protocol involving collagenase treatment of the worms is necessary for some antisera (e.g. anti-serotonin). Dauers are not permeabilized by this protocol and thus don't sta...
Monday, May 03, 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 1. Dissolve 2mg of IgG in 1mL of PBS...
Monday, May 03, 2004 Description Affinity Purification of Antibodies Using Proteins Bound to Nitrocellulose Procedure 1. Blot proteins onto nitrocellulose by standard electrophoretic transfer, visualize the proteins by Ponceau S staining,...
Monday, May 03, 2004 Description Antibody purification is possible because the cepharose beads are conjugated with protein which has an affinity under certain conditions to immunoglobulins. Following, you'll find a basic protocol for antib...
Monday, May 03, 2004 Description The following method is recommended for purification of monoclonal antibodies produced by tissue culture (hollow fibre cartridges or roller cultures) for laboratory use. Procedure General notes: 1) Endotoxi...