Histochemical Staining Protocol Protocol for histochemical staining. This protocol has been optimized for b-galactosidase and human placental alkaline phosphatase staining in retinal tissue and cultured cells.
Immunocytochemistry in Free-Floating Sections Protocol There are several strategies to visualize the antibody. For transmitted light microscopy, color development substrates for enzymes are often used. The antibody can be directly
labeled with the enzyme. However, such a covalent link between an antibody and an enzyme might result in a loss of both enzyme and antibody activity. For these reasons
several multistep staining procedures have been developed, where intermediate link antibodies are used. In this protocol use the Vectastain ABC-kit.
Preparing Frozen Tissue Sections for Immunostaining Protocol Frozen tissue sections show good preservation of tissue structure and antigens. The principle disadvantages of using them in immunostaining are that the specimens must be stored frozen, and a special microtome, known as a cryostat, is required. Also, many clinical specimens are not available in this form, and most classic histological descriptions of tissue structure and pathology are based on the use of paraffin-embedded sections of formalin-fixed material.
Preparing Paraffin Tissue Sections for Immunostaining Protocol Most histological studies are carried out on paraformaldehyde-fixed, paraffin-embedded tissue samples. Therefore, there is an extensive atlas of most tissues and organs prepared from these sources, and comparing the location of antigens to these data is immediately informative. The fixation and embedding procedures are harsh, however, and many antigens are not well preserved.
RUVKUN Antibody Staining Protocol Protocol for RUVKUN antibody staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING.