assaying Protocols

Category: Protein Protocols: Protein Ubiquitination Protocols

A Method for Assaying Deubiquitinating Enzymes. Lee et al., Biol Proced Online. May 1998. A general method for the assay of deubiquitinating enzymes was described in detail using 125I-labeled ubiquitin-fused αNH-MHISPPEPESEEEEEHYC (referred to as Ub-PESTc) as a substrate. - [Read A Method for Assaying Deubiquitinating Enzymes]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes the DNA content of bacteriophage lambda stocks can be easily and rapidly estimated. - [Read Assaying the DNA Content of Bacteriophage lambda Stocks and Lysates by Gel Electrophoresis Protocol]

Category: Immunology: B Cell Protocols

B cell activation can be quantitated indirectly by assaying antibody production or directly by measuring cellular changes that occur immediately after exposure to an activation signal. Provides methods for the latter (direct) approach--namely, methods for quantifying early parameters of B cell activation such as increases in intracellular ionized calcium concentration [Ca2+]I, cell size, and MHC class II-antigen expression. - [Read Early Events in B Lymphocyte Activation Protocol]

Category: Signalling Signal Transduction Protocols

This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography. Includes information: H1 Kinase Assay on Individual Xenopus Oocytes; H1 Kinase Assay on Xenopus Egg Extract Samples; H1 Kinase Assay on Tissue Culture Cells; Helpful protocol hints. - [Read Histone H1 Kinase Activity Assay]

Category: Fungi Protocols: Aspergillus Protocols

Gliotoxin is a metabolite of Aspergillus fumigatus that exhibits immunosuppressive activity against certain cells of the immune system. Secretion of gliotoxin during infection has been suggested as being a factor in the pathogenesis of aspergillosis. Gliotoxin secretion can be assayed in a number of ways by thin layer chromatography (TLC) high performance liquid chromatography (HPLC) or bioassay using the effect of gliotoxin on human cells1. - [Read Method for Assaying Gliotoxin Production in Aspergillus fumigatus Protocol]

Category: Antibody Protocols: Antibody Production Protocols

Monoclonal Antibody Production. Includes: Mouse Immunization; Bleeding Mice; Hybridoma Fusion; Assaying for Positive Clones; Expanding Positive Clones. - [Read Monoclonal Antibody Production]

Category: Cell Biology and Cell Culture

This protocol introduces the use of a liquid-filled wash chamber that separates unbound cells by gravity thereby eliminating uncontrolled shear forces and passage of adherent cells through a liquid/air interface. The cells are loaded with a fluorescent dye (6-carboxyfluorescein diacetate) for detection although other methods such as radioactive labels malabels may be used. This protocol is also useful for assaying molecules that promote or inhibit cell adhesion. - [Read Protocol for Measurement of Cell Adhesion Under Static Conditions]