Saturday, October 18, 2003 Description A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surface molecules and intracellular cytokines at single-cell...

Saturday, October 18, 2003 Description Matirgel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens, laminin, and proteoglycans)but also matrix degrading enz...

Saturday, October 18, 2003 Description This procedure allows detection of specific cell adhesion to glycolipids resolved on TLC plates. The steps include: (1) HPTLC of glycoconjugates, (2) coating the chromatogram with polymer, (3) prebloc...

Sunday, October 19, 2003 Description This protocol can be used to measure the amount of incorporation of 35S-Cysteine into cellular protein following a metabolic labeling procedure. This protocol reduces the amount of non-specific counts,...

Sunday, October 19, 2003 Description This protocol describes how to perform a DEAE-dextran transfection of COS cells and provides information on labeling the expressed protein with [35S-Cysteine] for SDS-PAGE analysis. Procedure A. Transfe...

Monday, October 20, 2003 Description This protocol describes an RNA Polymerase III (Pol III) transcription assay using an extract or proteins of choice. Pol III is the polymerase responsible for transcribing 5S RNA, tRNAs, and other small...

Monday, October 20, 2003 Description This protocol produces RNA transcripts from a cloned gene that can then be used in an in vitro translation assay. Procedure 1. Digest the gene-containing plasmid DNA to completion with a restriction end...

Monday, October 20, 2003 Description All reagents and tubes should be RNase-free. Make solutions up in plastic vessels using ddH2O. Autoclave the NaCl, MgCl2, Tris, glycerol, and gelatin solutions. Do not touch the undersides of tube c...

Tuesday, October 21, 2003 Description Labeling Oligonucleotides Procedure 1. Combine the following reactants in a microcentrifuge tube: 0.5 靗 of Denaturing Buffer 50 ng of the Oligonucleotide to be labeled (in TE Buffer) Bring the total r...

Tuesday, October 21, 2003 Description This assay uses a simple apparatus to study chemotaxis of leukocytes or other migratory cells. The apparatus consists of two multi-well chambers separated by a filter containing pores of uniform size....